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Showing 1 - 50 of 797
Showing 1 - 50 of 797
Details | EV-TRACK ID | Experiment nr. | Species | Sample type | Separation protocol | First author | Year | EV-METRIC |
---|---|---|---|---|---|---|---|---|
EV200179 | 3/4 | Homo sapiens | Primary human trophoblast |
(d)(U)C Filtration DG UF |
Ouyang, Yingshi | 2016 | 88% | |
Study summaryFull title
All authors
Yingshi Ouyang, Avraham Bayer, Tianjiao Chu, Vladimir A Tyurin, Valerian E Kagan, Adrian E Morelli, Carolyn B Coyne, Yoel Sadovsky
Journal
Placenta
Abstract
Introduction: Primary human trophoblasts release a repertoire of extracellular vesicles (EVs). Among (show more...)
EV-METRIC
88% (98th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Cell culture supernatant
Sample origin
Control condition
Focus vesicles
exosome
Separation protocol
Separation protocol
(Differential) (ultra)centrifugation
Filtration Density gradient Ultrafiltration Protein markers
EV: TSG101/ CD63/ Syntenin-1
non-EV: Argonaute2 Proteomics
yes
EV density (g/ml)
1.08-1.10
Show all info
Study aim
Function/Biomarker/Identification of content (omics approaches)
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
EV-producing cells
Primary human trophoblast
EV-harvesting Medium
EV-depleted medium
Preparation of EDS
Overnight, 100000g or commercial
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 100,000 g and 150,000 g
Pelleting performed
No
Density gradient
Type
Continuous
Lowest density fraction
6
Highest density fraction
40
Total gradient volume, incl. sample (mL)
12
Sample volume (mL)
1.5
Orientation
Bottom-up
Rotor type
Not specified
Speed (g)
100000
Duration (min)
Overnight
Fraction volume (mL)
.
Fraction processing
Ultrafiltration
Filtration steps
0.22µm or 0.2µm
Ultra filtration
Cut-off size (kDa)
100 kda
Membrane type
PES
Characterization: Protein analysis
Protein Concentration Method
BCA
Western Blot
Detected EV-associated proteins
CD63/ Syntenin-1/ TSG101
Not detected contaminants
Argonaute2
Proteomics database
Yes: Data and Specimen Hub
Characterization: RNA analysis
RNA analysis
Type
(RT)(q)PCR
Database
No
Proteinase treatment
No
RNAse treatment
No
Characterization: Lipid analysis
Yes
Characterization: Particle analysis
NTA
Report type
Mode
Reported size (nm)
80-90nm
EV concentration
Yes
EM
EM-type
Transmission-EM
Image type
Close-up
|
||||||||
EV160001 | 1/1 | Mus musculus | B16F10, JAWSII |
DG (d)(U)C |
Stremersch S | 2016 | 87% | |
Study summaryFull title
All authors
Stremersch S, Vandenbroucke RE, Van Wonterghem E, Hendrix A, De Smedt SC, Raemdonck K
Journal
J Control Release
Abstract
Exosome-like vesicles (ELVs) play an important role in intercellular communication by acting as natu (show more...)
EV-METRIC
87% (98th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Cell culture supernatant
Sample origin
Control condition
Focus vesicles
exosome-like vesicles
Separation protocol
Separation protocol
DG
(d)(U)C Protein markers
EV: CD81/ HSP70/ beta-actin/ CD63
non-EV: GM130/ Calreticulin Proteomics
no
Show all info
Study aim
Function, Mechanism of uptake/transfer
Sample
Species
Mus musculus
Sample Type
Cell culture supernatant
EV-producing cells
B16F10, JAWSII
EV-harvesting Medium
EV-depleted serum
Preparation of EDS
Ultrafiltration (MWCO 300 kDa)
Cell viability (%)
NA
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Below or equal to 800 g
Between 10,000 g and 50,000 g Pelleting performed
No
Density gradient
Only used for validation of main results
Yes
Density medium
115.5
Type
Discontinuous
Number of initial discontinuous layers
5
Lowest density fraction
0.125
Highest density fraction
0.5
Sample volume (mL)
1
Orientation
Top-down (sample migrates downwards)
Rotor type
SW 55 Ti
Speed (g)
200000
Duration (min)
900
Fraction volume (mL)
1
Fraction processing
Centrifugation
Pelleting: volume per fraction
5
Pelleting: duration (min)
150
Pelleting: rotor type
SW 55 Ti
Pelleting: speed (g)
120000
Pelleting: adjusted k-factor
115.5
Pelleting-wash: volume per pellet (mL)
5
Pelleting-wash: duration (min)
70
Pelleting-wash: rotor type
115.5
Pelleting-wash: speed (g)
SW 55 Ti
Pelleting-wash: adjusted k-factor
115.5
EV-subtype
Used subtypes
NO
Characterization: Protein analysis
Protein Concentration Method
BCA
Western Blot
Detected EV-associated proteins
CD63, CD81, HSP70, beta-actin
Flow cytometry specific beads
Selected surface protein(s)
CD63
Characterization: RNA analysis
Database
No
Proteinase treatment
No
RNAse treatment
No
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Size range/distribution
Reported size (nm)
30-300
EV concentration
Yes
Particle yield
1000
EM
EM-type
Cryo-EM
Image type
Close-up
|
||||||||
EV220152 | 3/6 | Mus musculus | 67NR |
(d)(U)C DG |
Vardaki I | 2016 | 78% | |
Study summaryFull title
All authors
Vardaki I, Ceder S, Rutishauser D, Baltatzis G, Foukakis T, Panaretakis T
Journal
Oncotarget
Abstract
Breast cancer (BrCa) is the most frequent cancer type in women and a leading cause of cancer related (show more...)
EV-METRIC
78% (97th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Cell culture supernatant
Sample origin
Control condition
Focus vesicles
exosome
Separation protocol
Separation protocol
(Differential) (ultra)centrifugation
Density gradient Protein markers
EV: CD63/ TSG101/ Rab5/ integrin alpha2/ integrin beta1/ periostin/ N-cadherin/ LOXL3/ LOXL4/ Glypican-1/ Glypican-4/ V-ATPase/ Alix/ beta-catenin/ E-cadherin/ Syndecan-4/ Vimentin/ GADPH/ AIF
non-EV: None Proteomics
yes
EV density (g/ml)
1.12-1.16
Show all info
Study aim
Biomarker/Identification of content (omics approaches)
Sample
Species
Mus musculus
Sample Type
Cell culture supernatant
EV-producing cells
67NR
EV-harvesting Medium
EV-depleted medium
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 800 g and 10,000 g
Between 100,000 g and 150,000 g Pelleting performed
Yes
Pelleting: speed (g)
120000
Wash: time (min)
120
Wash: speed (g)
120000
Density gradient
Only used for validation of main results
Yes
Lowest density fraction
0.2 M
Highest density fraction
2 M
Orientation
Top-down
Speed (g)
120000
Duration (min)
1200
Fraction processing
Centrifugation
Pelleting: duration (min)
120
Pelleting: speed (g)
120000
Characterization: Protein analysis
Protein Concentration Method
Bradford
Western Blot
Detected EV-associated proteins
CD63/ TSG101/ Rab5/ integrin alpha2/ integrin beta1/ periostin/ N-cadherin/ LOXL3/ LOXL4/ Glypican-1/ Glypican-4/ V-ATPase/ Alix
Not detected EV-associated proteins
beta-catenin/ E-cadherin/ Syndecan-4/ Vimentin/ GADPH/ AIF
Flow cytometry aspecific beads
Flow cytometry specific beads
Detected EV-associated proteins
CD63
Proteomics database
No
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Mean
Reported size (nm)
89
EV concentration
Yes
EM
EM-type
Transmission-EM
Image type
Close-up, Wide-field
|
||||||||
EV220152 | 4/6 | Mus musculus | 4T1 |
(d)(U)C DG |
Vardaki I | 2016 | 78% | |
Study summaryFull title
All authors
Vardaki I, Ceder S, Rutishauser D, Baltatzis G, Foukakis T, Panaretakis T
Journal
Oncotarget
Abstract
Breast cancer (BrCa) is the most frequent cancer type in women and a leading cause of cancer related (show more...)
EV-METRIC
78% (97th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Cell culture supernatant
Sample origin
Control condition
Focus vesicles
exosome
Separation protocol
Separation protocol
(Differential) (ultra)centrifugation
Density gradient Protein markers
EV: CD63/ TSG101/ Rab5/ Alix/ beta-catenin/ integrin alpha2/ integrin beta1/ periostin/ E-cadherin/ LOXL4/ Glypican-1/ V-ATPase/ Alix/ Vimentin/ GAPDH/ AIF/ N-Cadherin/ LOXL3/ Syndecan-4/ Glypican-4/ LOXL3/ N-Cadherin
non-EV: None Proteomics
yes
EV density (g/ml)
1.12-1.16
Show all info
Study aim
Biomarker/Identification of content (omics approaches)
Sample
Species
Mus musculus
Sample Type
Cell culture supernatant
EV-producing cells
4T1
EV-harvesting Medium
EV-depleted medium
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 800 g and 10,000 g
Between 100,000 g and 150,000 g Pelleting performed
Yes
Pelleting: speed (g)
120000
Wash: time (min)
120
Wash: speed (g)
120000
Density gradient
Only used for validation of main results
Yes
Lowest density fraction
0.2 M
Highest density fraction
2 M
Orientation
Top-down
Speed (g)
120000
Duration (min)
1200
Fraction processing
Centrifugation
Pelleting: duration (min)
120
Pelleting: speed (g)
120000
Characterization: Protein analysis
Protein Concentration Method
Bradford
Western Blot
Detected EV-associated proteins
CD63/ TSG101/ Rab5/ Alix/ beta-catenin/ integrin alpha2/ integrin beta1/ periostin/ E-cadherin/ LOXL4/ Glypican-1/ V-ATPase/ Alix
Not detected EV-associated proteins
Vimentin/ GAPDH/ AIF/ N-Cadherin/ LOXL3/ Syndecan-4/ Glypican-4/ LOXL3/ N-Cadherin
Flow cytometry aspecific beads
Flow cytometry specific beads
Detected EV-associated proteins
CD81
Proteomics database
No
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Mean
Reported size (nm)
102
EV concentration
Yes
EM
EM-type
Transmission-EM
Image type
Close-up, Wide-field
|
||||||||
EV210135 | 1/2 | Homo sapiens | Red blood cells |
DG (d)(U)C UF |
Stremersch, Stephan | 2016 | 78% | |
Study summaryFull title
All authors
Stephan Stremersch, Monica Marro, Bat-El Pinchasik, Pieter Baatsen, An Hendrix, Stefaan C De Smedt, Pablo Loza-Alvarez, Andre G Skirtach, Koen Raemdonck, Kevin Braeckmans
Journal
Small
Abstract
Exosome-like vesicles (ELVs) are a novel class of biomarkers that are receiving a lot of attention f (show more...)
EV-METRIC
78% (97th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Cell culture supernatant
Sample origin
Control condition
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
DG
(d)(U)C UF Protein markers
EV: CD81/ HSP70/ CD63/ actin
non-EV: None Proteomics
no
EV density (g/ml)
1.14
Show all info
Study aim
Biomarker/Technical analysis comparing/optimizing EV-related methods/New methodological development
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
EV-producing cells
Red blood cells
EV-harvesting Medium
Serum free medium
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Below or equal to 800 g
Between 800 g and 10,000 g Between 10,000 g and 50,000 g Pelleting performed
No
Density gradient
Type
Discontinuous
Number of initial discontinuous layers
4
Lowest density fraction
12.5%
Highest density fraction
50%
Total gradient volume, incl. sample (mL)
Not specified
Sample volume (mL)
Not spec
Orientation
Top-down
Rotor type
SW 55 Ti
Speed (g)
150000
Duration (min)
900
Fraction volume (mL)
0.5
Fraction processing
Centrifugation
Pelleting: volume per fraction
5
Pelleting: duration (min)
150
Pelleting: rotor type
SW 55 Ti
Pelleting: speed (g)
150000
Ultra filtration
Cut-off size (kDa)
30
Membrane type
Not specified
Characterization: Protein analysis
Protein Concentration Method
BCA
Western Blot
Detected EV-associated proteins
CD63/ actin/ HSP70/ CD81
Characterization: Lipid analysis
Yes
Characterization: Particle analysis
NTA
Report type
Modus
Reported size (nm)
170
Particle yield
Not reported
EM
EM-type
Cryo-EM
Image type
Wide-field
EV concentration
Not
|
||||||||
EV210135 | 2/2 | Mus musculus | B16F10 |
DG (d)(U)C UF |
Stremersch, Stephan | 2016 | 78% | |
Study summaryFull title
All authors
Stephan Stremersch, Monica Marro, Bat-El Pinchasik, Pieter Baatsen, An Hendrix, Stefaan C De Smedt, Pablo Loza-Alvarez, Andre G Skirtach, Koen Raemdonck, Kevin Braeckmans
Journal
Small
Abstract
Exosome-like vesicles (ELVs) are a novel class of biomarkers that are receiving a lot of attention f (show more...)
EV-METRIC
78% (97th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Cell culture supernatant
Sample origin
Control condition
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
DG
(d)(U)C UF Protein markers
EV: CD81/ HSP70/ CD63/ actin
non-EV: None Proteomics
no
EV density (g/ml)
1.14
Show all info
Study aim
Biomarker/Technical analysis comparing/optimizing EV-related methods/New methodological development
Sample
Species
Mus musculus
Sample Type
Cell culture supernatant
EV-producing cells
B16F10
EV-harvesting Medium
EV-depleted medium
Preparation of EDS
Other preparation;Ultrafiltration
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Below or equal to 800 g
Between 800 g and 10,000 g Between 10,000 g and 50,000 g Pelleting performed
No
Density gradient
Type
Discontinuous
Number of initial discontinuous layers
4
Lowest density fraction
12.5%
Highest density fraction
50%
Total gradient volume, incl. sample (mL)
Not specified
Sample volume (mL)
Not spec
Orientation
Top-down
Rotor type
SW 55 Ti
Speed (g)
150000
Duration (min)
900
Fraction volume (mL)
0.5
Fraction processing
Centrifugation
Pelleting: volume per fraction
5
Pelleting: duration (min)
150
Pelleting: rotor type
SW 55 Ti
Pelleting: speed (g)
150000
Ultra filtration
Cut-off size (kDa)
30
Membrane type
Not specified
Characterization: Protein analysis
Protein Concentration Method
BCA
Western Blot
Detected EV-associated proteins
CD63/ actin/ HSP70/ CD81
Characterization: Lipid analysis
Yes
Characterization: Particle analysis
NTA
Report type
Modus
Reported size (nm)
120
Particle yield
Not reported
EM
EM-type
Cryo-EM
Image type
Wide-field
EV concentration
Not
|
||||||||
EV210035 | 2/2 | Homo sapiens | A-431 |
(d)(U)C DG Filtration |
van Dommelen, Susan M | 2016 | 78% | |
Study summaryFull title
All authors
Susan M van Dommelen, Roy van der Meel, Wouter W van Solinge, Maria Coimbra, Pieter Vader, Raymond M Schiffelers
Journal
Nanomedicine
Abstract
Aim: Extracellular vesicles (EVs) are attractive candidates for biomarker research, because their co (show more...)
EV-METRIC
78% (97th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Cell culture supernatant
Sample origin
Cetuximab
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
(Differential) (ultra)centrifugation
Density gradient Filtration Protein markers
EV: TSG101/ Alix/ CD9/ EGFR/ pEGFR/ Akt/ pAkt/ -actin/ Cetuximab
non-EV: Lamin-A/ Lamin-C/ ATP5-A/ Tom20 Proteomics
no
EV density (g/ml)
1.10-1.21
Show all info
Study aim
Function/Identification of content (omics approaches)
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
EV-producing cells
A-431
EV-harvesting Medium
Serum free medium
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Below or equal to 800 g
Between 800 g and 10,000 g Between 100,000 g and 150,000 g Pelleting performed
Yes
Pelleting: time(min)
70
Pelleting: rotor type
JA-30.50
Pelleting: speed (g)
100000
Wash: volume per pellet (ml)
Not specified
Wash: time (min)
70
Wash: Rotor Type
JA-30.50
Wash: speed (g)
100000
Density gradient
Only used for validation of main results
Yes
Type
Continuous
Lowest density fraction
0.4
Highest density fraction
2.5
Total gradient volume, incl. sample (mL)
Not specified
Sample volume (mL)
Not spec
Orientation
Top-down
Rotor type
SW 40 Ti
Speed (g)
200000
Duration (min)
960
Fraction volume (mL)
1
Fraction processing
Centrifugation
Pelleting: volume per fraction
4
Pelleting: duration (min)
70
Pelleting: rotor type
Not specified
Pelleting: speed (g)
100000
Filtration steps
0.22µm or 0.2µm
Characterization: Protein analysis
Protein Concentration Method
microBCA
Western Blot
Detected EV-associated proteins
CD9/ EGFR/ pEGFR/ Akt/ pAkt/ -actin/ Cetuximab/ TSG101/ Alix
Not detected contaminants
Lamin-A/ Lamin-C/ ATP5-A/ Tom20
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Not Reported
EV concentration
Yes
|
||||||||
EV200179 | 4/4 | Homo sapiens | Blood plasma |
(d)(U)C Filtration DG UF PEG precipitaton Gelatin-sepharose chromatograhy |
Ouyang, Yingshi | 2016 | 75% | |
Study summaryFull title
All authors
Yingshi Ouyang, Avraham Bayer, Tianjiao Chu, Vladimir A Tyurin, Valerian E Kagan, Adrian E Morelli, Carolyn B Coyne, Yoel Sadovsky
Journal
Placenta
Abstract
Introduction: Primary human trophoblasts release a repertoire of extracellular vesicles (EVs). Among (show more...)
EV-METRIC
75% (96th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Blood plasma
Sample origin
Healthy pregnant
Focus vesicles
exosome
Separation protocol
Separation protocol
(Differential) (ultra)centrifugation
Filtration Density gradient Ultrafiltration PEG precipitaton Gelatin-sepharose chromatograhy Protein markers
EV: CD63
non-EV: Fibronectin Proteomics
yes
EV density (g/ml)
1.08-1.10
Show all info
Study aim
Function/Biomarker/Identification of content (omics approaches)
Sample
Species
Homo sapiens
Sample Type
Blood plasma
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 800 g and 10,000 g
Pelleting performed
No
Density gradient
Type
Continuous
Lowest density fraction
6
Highest density fraction
30
Total gradient volume, incl. sample (mL)
14
Sample volume (mL)
0.5
Orientation
Top-down
Rotor type
Not specified
Speed (g)
10000
Duration (min)
Overnight
Fraction volume (mL)
.
Fraction processing
Ultrafiltration
Pelleting: volume per fraction
.
Filtration steps
0.22µm or 0.2µm
Ultra filtration
Cut-off size (kDa)
100 kda
Membrane type
PES
Other
Name other separation method
PEG precipitaton
Other
Name other separation method
Gelatin-sepharose chromatograhy
Characterization: Protein analysis
Protein Concentration Method
BCA
Western Blot
Detected EV-associated proteins
CD63
Not detected contaminants
Fibronectin
Proteomics database
No
Characterization: RNA analysis
RNA analysis
Type
(RT)(q)PCR
Database
No
Proteinase treatment
No
RNAse treatment
No
Characterization: Lipid analysis
Yes
Characterization: Particle analysis
NTA
Report type
Mode
Reported size (nm)
80-90nm
EV concentration
Yes
|
||||||||
EV230697 | 1/3 | Porphyromonas gingivalis | W50 |
(d)(U)C DG Filtration UF |
Cecil JD | 2016 | 67% | |
Study summaryFull title
All authors
Cecil JD, O'Brien-Simpson NM, Lenzo JC, Holden JA, Chen YY, Singleton W, Gause KT, Yan Y, Caruso F, Reynolds EC
Journal
PLoS One
Abstract
Highly purified outer membrane vesicles (OMVs) of the periodontal pathogens, Porphyromonas gingivali (show more...)
EV-METRIC
67% (94th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Cell culture supernatant
Sample origin
Control condition
Focus vesicles
outer membrane vesicles
Separation protocol
Separation protocol
(Differential) (ultra)centrifugation
Density gradient Filtration Ultrafiltration Protein markers
EV: None
non-EV: None Proteomics
no
EV density (g/ml)
not specified
Show all info
Study aim
Function
Sample
Species
Porphyromonas gingivalis
Sample Type
Cell culture supernatant
EV-producing cells
W50
EV-harvesting Medium
Serum-containing medium
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 800 g and 10,000 g
Between 100,000 g and 150,000 g Pelleting performed
Yes
Pelleting: rotor type
JA-30.50 Ti
Pelleting: speed (g)
100000
Density gradient
Type
Discontinuous
Number of initial discontinuous layers
7
Lowest density fraction
15%
Highest density fraction
45%
Total gradient volume, incl. sample (mL)
12
Sample volume (mL)
not spec
Orientation
Bottom-up
Speed (g)
150000
Duration (min)
960
Fraction volume (mL)
1.5
Fraction processing
Centrifugation
Pelleting: duration (min)
120
Pelleting: rotor type
SW 40 Ti
Pelleting: speed (g)
150000
Filtration steps
0.2 or 0.22 µm
Ultra filtration
Cut-off size (kDa)
100
Membrane type
Polyethersulfone (PES)
Characterization: Protein analysis
None
Protein Concentration Method
Fluorometric assay
Characterization: RNA analysis
RNA analysis
Type
Qubit RNA assay kit
Proteinase treatment
No
RNAse treatment
No
Characterization: Lipid analysis
Yes
Characterization: Particle analysis
DLS
Report type
Mean
Reported size (nm)
121
Particle analysis: flow cytometry
Flow cytometer type
Apogee A50-Micro Flow Cytometer
Hardware adjustment
Calibration bead size
0.11
EV concentration
Yes
Particle yield
particles per milliliter of starting sample: 4.98E+12
EM
EM-type
Cryo-EM
Image type
Close-up, Wide-field
Report size (nm)
50-150
|
||||||||
EV230697 | 2/3 | Treponema denticola | ATCC 35405 |
(d)(U)C DG Filtration UF |
Cecil JD | 2016 | 67% | |
Study summaryFull title
All authors
Cecil JD, O'Brien-Simpson NM, Lenzo JC, Holden JA, Chen YY, Singleton W, Gause KT, Yan Y, Caruso F, Reynolds EC
Journal
PLoS One
Abstract
Highly purified outer membrane vesicles (OMVs) of the periodontal pathogens, Porphyromonas gingivali (show more...)
EV-METRIC
67% (94th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Cell culture supernatant
Sample origin
Control condition
Focus vesicles
outer membrane vesicles
Separation protocol
Separation protocol
(Differential) (ultra)centrifugation
Density gradient Filtration Ultrafiltration Protein markers
EV: None
non-EV: None Proteomics
no
EV density (g/ml)
not specified
Show all info
Study aim
Function
Sample
Species
Treponema denticola
Sample Type
Cell culture supernatant
EV-producing cells
ATCC 35405
EV-harvesting Medium
Serum-containing medium
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 800 g and 10,000 g
Between 100,000 g and 150,000 g Pelleting performed
Yes
Pelleting: rotor type
JA-30.50 Ti
Pelleting: speed (g)
100000
Density gradient
Type
Discontinuous
Number of initial discontinuous layers
7
Lowest density fraction
15%
Highest density fraction
45%
Total gradient volume, incl. sample (mL)
12
Sample volume (mL)
not spec
Orientation
Bottom-up
Speed (g)
150000
Duration (min)
2880
Fraction volume (mL)
1.5
Fraction processing
Centrifugation
Pelleting: duration (min)
120
Pelleting: rotor type
SW 40 Ti
Pelleting: speed (g)
150000
Filtration steps
0.2 or 0.22 µm
Ultra filtration
Cut-off size (kDa)
100
Membrane type
Polyethersulfone (PES)
Characterization: Protein analysis
None
Protein Concentration Method
Fluorometric assay
Characterization: RNA analysis
RNA analysis
Type
Qubit RNA assay kit
Proteinase treatment
No
RNAse treatment
No
Characterization: Lipid analysis
Yes
Characterization: Particle analysis
DLS
Report type
Mean
Reported size (nm)
75
Particle analysis: flow cytometry
Flow cytometer type
Apogee A50-Micro Flow Cytometer
Hardware adjustment
Calibration bead size
0.11
EV concentration
Yes
Particle yield
particles per milliliter of starting sample: 5.54E+11
EM
EM-type
Cryo-EM
Image type
Close-up, Wide-field
Report size (nm)
30-120
|
||||||||
EV230697 | 3/3 | Tannerella forsythia | ATCC 43037 |
(d)(U)C DG Filtration UF |
Cecil JD | 2016 | 67% | |
Study summaryFull title
All authors
Cecil JD, O'Brien-Simpson NM, Lenzo JC, Holden JA, Chen YY, Singleton W, Gause KT, Yan Y, Caruso F, Reynolds EC
Journal
PLoS One
Abstract
Highly purified outer membrane vesicles (OMVs) of the periodontal pathogens, Porphyromonas gingivali (show more...)
EV-METRIC
67% (94th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Cell culture supernatant
Sample origin
Control condition
Focus vesicles
outer membrane vesicles
Separation protocol
Separation protocol
(Differential) (ultra)centrifugation
Density gradient Filtration Ultrafiltration Protein markers
EV: None
non-EV: None Proteomics
no
EV density (g/ml)
not specified
Show all info
Study aim
Function
Sample
Species
Tannerella forsythia
Sample Type
Cell culture supernatant
EV-producing cells
ATCC 43037
EV-harvesting Medium
Serum-containing medium
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 800 g and 10,000 g
Between 100,000 g and 150,000 g Pelleting performed
Yes
Pelleting: rotor type
JA-30.50 Ti
Pelleting: speed (g)
100000
Density gradient
Type
Discontinuous
Number of initial discontinuous layers
7
Lowest density fraction
15%
Highest density fraction
45%
Total gradient volume, incl. sample (mL)
12
Sample volume (mL)
not spec
Orientation
Bottom-up
Speed (g)
150000
Duration (min)
2880
Fraction volume (mL)
1.5
Fraction processing
Centrifugation
Pelleting: duration (min)
120
Pelleting: rotor type
SW 40 Ti
Pelleting: speed (g)
150000
Filtration steps
0.2 or 0.22 µm
Ultra filtration
Cut-off size (kDa)
100
Membrane type
Polyethersulfone (PES)
Characterization: Protein analysis
None
Protein Concentration Method
Fluorometric assay
Characterization: RNA analysis
RNA analysis
Type
Qubit RNA assay kit
Proteinase treatment
No
RNAse treatment
No
Characterization: Lipid analysis
Yes
Characterization: Particle analysis
DLS
Report type
Mean
Reported size (nm)
158
Particle analysis: flow cytometry
Flow cytometer type
Apogee A50-Micro Flow Cytometer
Hardware adjustment
Calibration bead size
0.11
EV concentration
Yes
Particle yield
particles per milliliter of starting sample: 2.00E+12
EM
EM-type
Cryo-EM
Image type
Close-up, Wide-field
Report size (nm)
80-250
|
||||||||
EV210487 | 1/2 | Mus musculus | MIN6 |
DG (d)(U)C UF Filtration |
Bosch S | 2016 | 67% | |
Study summaryFull title
All authors
Bosch S, de Beaurepaire L, Allard M, Mosser M, Heichette C, Chrétien D, Jegou D, Bach JM
Journal
Sci Rep
Abstract
Exosomes are important mediators in intercellular communication. Released by many cell types, they t (show more...)
EV-METRIC
67% (94th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Cell culture supernatant
Sample origin
Control condition
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
Density gradient
(Differential) (ultra)centrifugation Ultrafiltration Filtration Protein markers
EV: CD81/ CD63
non-EV: Calnexin Proteomics
no
Show all info
Study aim
Technical analysis comparing/optimizing EV-related methods
Sample
Species
Mus musculus
Sample Type
Cell culture supernatant
EV-producing cells
MIN6
EV-harvesting Medium
EV-depleted medium
Preparation of EDS
overnight (16h) at >=100,000g
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Below or equal to 800 g
Between 100,000 g and 150,000 g Pelleting performed
Yes
Pelleting: rotor type
SW 41 Ti
Pelleting: speed (g)
100000
Density gradient
Only used for validation of main results
Yes
Type
Discontinuous
Number of initial discontinuous layers
4
Lowest density fraction
5%
Highest density fraction
40%
Total gradient volume, incl. sample (mL)
16.1
Sample volume (mL)
0.60
Orientation
Top-down
Speed (g)
100000
Duration (min)
1080
Fraction volume (mL)
1
Fraction processing
Centrifugation
Pelleting: volume per fraction
1
Pelleting: duration (min)
180
Pelleting: speed (g)
100000
Filtration steps
0.22µm or 0.2µm
Ultra filtration
Cut-off size (kDa)
100
Membrane type
Regenerated cellulose
Characterization: Protein analysis
Protein Concentration Method
Bradford
Western Blot
Detected EV-associated proteins
CD63/ CD81
Not detected contaminants
Calnexin
Characterization: RNA analysis
RNA analysis
Type
Capillary electrophoresis (e.g. Bioanalyzer)
Database
No
Proteinase treatment
No
RNAse treatment
No
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Modus
Reported size (nm)
113
EV concentration
Yes
EM
EM-type
Cryo-EM
Image type
Close-up, Wide-field
|
||||||||
EV210487 | 2/2 | Mus musculus | MIN6 |
DG (d)(U)C UF Filtration |
Bosch S | 2016 | 67% | |
Study summaryFull title
All authors
Bosch S, de Beaurepaire L, Allard M, Mosser M, Heichette C, Chrétien D, Jegou D, Bach JM
Journal
Sci Rep
Abstract
Exosomes are important mediators in intercellular communication. Released by many cell types, they t (show more...)
EV-METRIC
67% (94th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Cell culture supernatant
Sample origin
Trehalose
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
Density gradient
(Differential) (ultra)centrifugation Ultrafiltration Filtration Protein markers
EV: CD81/ CD63
non-EV: Calnexin Proteomics
no
Show all info
Study aim
Technical analysis comparing/optimizing EV-related methods
Sample
Species
Mus musculus
Sample Type
Cell culture supernatant
EV-producing cells
MIN6
EV-harvesting Medium
EV-depleted medium
Preparation of EDS
overnight (16h) at >=100,000g
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Below or equal to 800 g
Between 100,000 g and 150,000 g Pelleting performed
Yes
Pelleting: rotor type
SW 41 Ti
Pelleting: speed (g)
100000
Density gradient
Only used for validation of main results
Yes
Type
Discontinuous
Number of initial discontinuous layers
4
Lowest density fraction
5%
Highest density fraction
40%
Total gradient volume, incl. sample (mL)
16.1
Sample volume (mL)
0.60
Orientation
Top-down
Speed (g)
100000
Duration (min)
1080
Fraction volume (mL)
1
Fraction processing
Centrifugation
Pelleting: volume per fraction
1
Pelleting: duration (min)
180
Pelleting: speed (g)
100000
Filtration steps
0.22µm or 0.2µm
Ultra filtration
Cut-off size (kDa)
100
Membrane type
Regenerated cellulose
Characterization: Protein analysis
Protein Concentration Method
Bradford
Western Blot
Detected EV-associated proteins
CD63/ CD81
Not detected contaminants
Calnexin
Characterization: RNA analysis
RNA analysis
Type
Capillary electrophoresis (e.g. Bioanalyzer)
Database
No
Proteinase treatment
No
RNAse treatment
No
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Modus
Reported size (nm)
111
EV concentration
Yes
EM
EM-type
Cryo-EM
Image type
Close-up, Wide-field
|
||||||||
EV210101 | 4/6 | Homo sapiens | Urine |
(d)(U)C Filtration SEC (non-commercial) |
Welton, Joanne Louise | 2016 | 67% | |
Study summaryFull title
All authors
Joanne Louise Welton, Paul Brennan, Mark Gurney, Jason Paul Webber, Lisa Kate Spary, David Gil Carton, Juan Manuel Falcón-Pérez, Sean Peter Walton, Malcolm David Mason, Zsuzsanna Tabi, Aled Clayton
Journal
J Extracell Vesicles
Abstract
Proteomics analysis of biofluid-derived vesicles holds enormous potential for discovering non-invasi (show more...)
EV-METRIC
67% (94th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Urine
Sample origin
Control condition
Focus vesicles
exosome
Separation protocol
Separation protocol
(d)(U)C
Filtration Size-exclusion chromatography (non-commercial) Protein markers
EV: Alix/ TSG101/ LAMP2A
non-EV: Albumin Proteomics
no
Show all info
Study aim
Identification of content (omics approaches)/Technical analysis comparing/optimizing EV-related methods
Sample
Species
Homo sapiens
Sample Type
Urine
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Below or equal to 800 g
Between 800 g and 10,000 g Equal to or above 150,000 g Pelleting performed
Yes
Pelleting: time(min)
120
Pelleting: rotor type
Type 70 Ti
Pelleting: speed (g)
200000
Filtration steps
0.22µm or 0.2µm
Size-exclusion chromatography
Total column volume (mL)
2.8
Sample volume/column (mL)
0.5
Resin type
Sepharose CL-2B
Characterization: Protein analysis
Protein Concentration Method
Other;Spectrophotometry
Western Blot
Detected EV-associated proteins
Alix/ LAMP2A/ TSG101
Not detected contaminants
Albumin
Detected EV-associated proteins
SOMAscan multiplex assay
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Mean
Reported size (nm)
117.75
EV concentration
Yes
Particle yield
As the number of particles per µg protein;Yes, other: 20000000000
EM
EM-type
Cryo-EM
Image type
Close-up
|
||||||||
EV200134 | 2/2 | Homo sapiens | Blood plasma |
DG (d)(U)C Filtration |
Salomon, Carlos | 2016 | 67% | |
Study summaryFull title
All authors
Carlos Salomon, Katherin Scholz-Romero, Suchismita Sarker, Emma Sweeney, Miharu Kobayashi, Paula Correa, Sherri Longo, Gregory Duncombe, Murray D Mitchell, Gregory E Rice, Sebastian E Illanes
Journal
Diabetes
Abstract
Although there is significant interest in elucidating the role of placenta-derived exosomes (PdEs) d (show more...)
EV-METRIC
67% (93rd percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Blood plasma
Sample origin
Gestational diabetes mellitus
Focus vesicles
exosome
Separation protocol
Separation protocol
Density gradient
(Differential) (ultra)centrifugation Filtration Protein markers
EV: PLAP/ CD63/ TSG101
non-EV: None Proteomics
no
EV density (g/ml)
1.122-1.156
Show all info
Study aim
Function
Sample
Species
Homo sapiens
Sample Type
Blood plasma
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 800 g and 10,000 g
Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Pelleting performed
Yes
Pelleting: time(min)
120
Pelleting: rotor type
T-8100
Pelleting: speed (g)
100000
Wash: volume per pellet (ml)
10
Wash: time (min)
120
Wash: Rotor Type
T-8100
Wash: speed (g)
100000
Density gradient
Type
Discontinuous
Number of initial discontinuous layers
4
Lowest density fraction
5%
Highest density fraction
40%
Total gradient volume, incl. sample (mL)
14.5mL
Sample volume (mL)
0.5mL
Orientation
Top-down
Rotor type
T-8100
Speed (g)
100000
Duration (min)
1200
Fraction processing
Centrifugation
Pelleting: duration (min)
120
Pelleting: rotor type
T-8100
Pelleting: speed (g)
100000
Filtration steps
0.22µm or 0.2µm
Characterization: Protein analysis
Protein Concentration Method
Not determined
Western Blot
Detected EV-associated proteins
CD63/ TSG101
ELISA
Detected EV-associated proteins
PLAP
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Mean
Reported size (nm)
100 - 108nm
EV concentration
Yes
EM
EM-type
Transmission-EM
Image type
Close-up, Wide-field
|
||||||||
EV160004 | 2/5 | Equus caballus | Synovial fluid |
DG (d)(U)C |
Boere J | 2016 | 66% | |
Study summaryFull title
All authors
Boere J, van de Lest CH, Libregts SF, Arkesteijn GJ, Geerts WJ, Nolte-'t Hoen EN, Malda J, van Weeren PR, Wauben MH
Journal
J Extracell Vesicles
Abstract
Extracellular vesicles (EVs) in synovial fluid (SF) are gaining increased recognition as important f (show more...)
EV-METRIC
66% (82nd percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Synovial fluid
Sample origin
Control condition
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
DG
(d)(U)C Adj. k-factor
71.08 (pelleting)
Protein markers
EV: Annexin-A1/ CD90/Thy1.1
non-EV: None Proteomics
no
Show all info
Study aim
Biomarker, New methodological development
Sample
Species
Equus caballus
Sample Type
Synovial fluid
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 800 g and 10,000 g
Between 10,000 g and 50,000 g Equal to or above 150,000 g Pelleting performed
Yes
Pelleting: time(min)
120
Pelleting: rotor type
MLS-50
Pelleting: speed (g)
200000
Pelleting: adjusted k-factor
71.08
Density gradient
Type
Discontinuous
Number of initial discontinuous layers
3
Lowest density fraction
0M
Highest density fraction
1.4M
Sample volume (mL)
1.5
Orientation
Bottom-up (sample migrates upwards)
Rotor type
MLS-50
Speed (g)
200000
Duration (min)
960
Fraction volume (mL)
1
Fraction processing
Centrifugation
Pelleting: volume per fraction
12
Pelleting: duration (min)
60
Pelleting: rotor type
SW 40 Ti
Pelleting: speed (g)
200000
Pelleting: adjusted k-factor
138.3
Characterization: Protein analysis
Protein Concentration Method
Not determined
Western Blot
Detected EV-associated proteins
Annexin-A1, CD90/Thy1.1
Characterization: Lipid analysis
No
Characterization: Particle analysis
EM
EM-type
Cryo-EM
Image type
Close-up, Wide-field
Report size (nm)
20-200
Extra information
Importantly, synovial fluid samples were pre-treated with hyaluronidase prior to ultracentrifugation.
|
||||||||
EV160004 | 5/5 | Equus caballus | Synovial fluid |
DG (d)(U)C |
Boere J | 2016 | 66% | |
Study summaryFull title
All authors
Boere J, van de Lest CH, Libregts SF, Arkesteijn GJ, Geerts WJ, Nolte-'t Hoen EN, Malda J, van Weeren PR, Wauben MH
Journal
J Extracell Vesicles
Abstract
Extracellular vesicles (EVs) in synovial fluid (SF) are gaining increased recognition as important f (show more...)
EV-METRIC
66% (82nd percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Synovial fluid
Sample origin
Control condition
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
DG
(d)(U)C Adj. k-factor
1421 (pelleting)
Protein markers
EV: Annexin-A1/ CD90/Thy1.1
non-EV: None Proteomics
no
Show all info
Study aim
Biomarker, New methodological development
Sample
Species
Equus caballus
Sample Type
Synovial fluid
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 800 g and 10,000 g
Between 10,000 g and 50,000 g Pelleting performed
Yes
Pelleting: time(min)
120
Pelleting: rotor type
MLS-50
Pelleting: speed (g)
10000
Pelleting: adjusted k-factor
1421.
Density gradient
Type
Discontinuous
Number of initial discontinuous layers
3
Lowest density fraction
0M
Highest density fraction
1.4M
Sample volume (mL)
1.5
Orientation
Bottom-up (sample migrates upwards)
Rotor type
MLS-50
Speed (g)
200000
Duration (min)
960
Fraction volume (mL)
1
Fraction processing
Centrifugation
Pelleting: volume per fraction
12
Pelleting: duration (min)
60
Pelleting: rotor type
SW 40 Ti
Pelleting: speed (g)
200000
Pelleting: adjusted k-factor
138.3
Characterization: Protein analysis
Protein Concentration Method
Not determined
Western Blot
Detected EV-associated proteins
Annexin-A1, CD90/Thy1.1
Characterization: Lipid analysis
No
Characterization: Particle analysis
EM
EM-type
Cryo-EM
Image type
Close-up, Wide-field
Report size (nm)
20-200
Extra information
Importantly, synovial fluid samples were pre-treated with hyaluronidase prior to ultracentrifugation.
|
||||||||
EV200044 | 1/1 | Homo sapiens | Blood plasma |
Precipitation DG |
Zhang YN | 2016 | 63% | |
Study summaryFull title
All authors
Zhang YN, Vernooij F, Ibrahim I, Ooi S, Gijsberts CM, Schoneveld AH, Sen KW, den Ruijter HM, Timmers L, Richards AM, Jong CT, Mazlan I, Wang JW, Lam CS, de Kleijn DP
Journal
PLoS One
Abstract
BACKGROUND:
SerpinF2, SerpinG1, CystatinC and CD14 are involved in inflammatory processes and plasma (show more...)
EV-METRIC
63% (91st percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Blood plasma
Sample origin
Healthy test subject
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
Precipitation
Density gradient Protein markers
EV: CD14/ SerpinC1/ SerpinG1/ CystC/ SerpinF2
non-EV: None Proteomics
no
EV density (g/ml)
1.02-1.17
Show all info
Study aim
Biomarker/New methodological development
Sample
Species
Homo sapiens
Sample Type
Blood plasma
Separation Method
Density gradient
Only used for validation of main results
Yes
Type
Discontinuous
Number of initial discontinuous layers
5
Lowest density fraction
5%
Highest density fraction
40%
Total gradient volume, incl. sample (mL)
10.5
Sample volume (mL)
0.5
Orientation
Top-down
Rotor type
SW 41 Ti
Speed (g)
200000
Duration (min)
1080
Fraction volume (mL)
1
Fraction processing
Centrifugation
Pelleting: volume per fraction
8
Pelleting: duration (min)
60
Pelleting: rotor type
SW 41 Ti
Pelleting: speed (g)
200000
Other
Name other separation method
Precipitation
Other
Name other separation method
Density gradient
Characterization: Protein analysis
Protein Concentration Method
Not determined
Western Blot
Detected EV-associated proteins
CD9
ELISA
Detected EV-associated proteins
CD14/ SerpinC1/ SerpinG1/ CystC/ SerpinF2
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Size range/distribution
Reported size (nm)
50-120
|
||||||||
EV230700 | 1/2 | Vibrio mediterranei | AK1 |
(d)(U)C DG Filtration UF |
Li J | 2016 | 57% | |
Study summaryFull title
All authors
Li J, Azam F, Zhang S
Journal
Environ Microbiol
Abstract
Production and release of outer-membrane vesicles (OMVs) is known in many bacteria including human p (show more...)
EV-METRIC
57% (92nd percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Cell culture supernatant
Sample origin
Grown at 20°C
Focus vesicles
outer membrane vesicles
Separation protocol
Separation protocol
(Differential) (ultra)centrifugation
Density gradient Filtration Ultrafiltration Protein markers
EV: None
non-EV: None Proteomics
yes
EV density (g/ml)
not specified
Show all info
Study aim
Function
Sample
Species
Vibrio mediterranei
Sample Type
Cell culture supernatant
EV-producing cells
AK1
EV-harvesting Medium
Serum free medium
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 10,000 g and 50,000 g
Between 100,000 g and 150,000 g Pelleting performed
Yes
Pelleting: time(min)
120
Pelleting: rotor type
P27A
Pelleting: speed (g)
100000
Density gradient
Type
Discontinuous
Number of initial discontinuous layers
9
Lowest density fraction
0%
Highest density fraction
45%
Total gradient volume, incl. sample (mL)
not specified
Sample volume (mL)
1.3
Orientation
Bottom-up
Speed (g)
111132
Duration (min)
360
Fraction volume (mL)
not specified
Fraction processing
Centrifugation
Pelleting: volume per fraction
not spec
Pelleting: rotor type
P90AT
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
TDB
Filtration steps
0.2 or 0.22 µm
Ultra filtration
Cut-off size (kDa)
100
Membrane type
Polyethersulfone (PES)
Characterization: Protein analysis
Protein Concentration Method
Bradford
Protein Yield (µg)
per 4 l of starting sample
Proteomics database
No
Characterization: Lipid analysis
No
Characterization: Particle analysis
EM
EM-type
Transmission-EM
Image type
Wide-field
Report size (nm)
20-120
|
||||||||
EV230700 | 2/2 | Vibrio mediterranei | AK1 |
(d)(U)C DG Filtration UF |
Li J | 2016 | 57% | |
Study summaryFull title
All authors
Li J, Azam F, Zhang S
Journal
Environ Microbiol
Abstract
Production and release of outer-membrane vesicles (OMVs) is known in many bacteria including human p (show more...)
EV-METRIC
57% (92nd percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Cell culture supernatant
Sample origin
Grown at 30°C
Focus vesicles
outer membrane vesicles
Separation protocol
Separation protocol
(Differential) (ultra)centrifugation
Density gradient Filtration Ultrafiltration Protein markers
EV: None
non-EV: None Proteomics
yes
EV density (g/ml)
not specified
Show all info
Study aim
Function
Sample
Species
Vibrio mediterranei
Sample Type
Cell culture supernatant
EV-producing cells
AK1
EV-harvesting Medium
Serum free medium
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 10,000 g and 50,000 g
Between 100,000 g and 150,000 g Pelleting performed
Yes
Pelleting: time(min)
120
Pelleting: rotor type
P27A
Pelleting: speed (g)
100000
Density gradient
Type
Discontinuous
Number of initial discontinuous layers
9
Lowest density fraction
0%
Highest density fraction
45%
Total gradient volume, incl. sample (mL)
not specified
Sample volume (mL)
1.3
Orientation
Bottom-up
Speed (g)
111132
Duration (min)
360
Fraction volume (mL)
not specified
Fraction processing
Centrifugation
Pelleting: volume per fraction
not spec
Pelleting: rotor type
P90AT
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
TDB
Filtration steps
0.2 or 0.22 µm
Ultra filtration
Cut-off size (kDa)
100
Membrane type
Polyethersulfone (PES)
Characterization: Protein analysis
Protein Concentration Method
Bradford
Protein Yield (µg)
per 4 l of starting sample
Proteomics database
No
Characterization: Lipid analysis
No
Characterization: Particle analysis
EM
EM-type
Transmission-EM
Image type
Wide-field
Report size (nm)
20-120
|
||||||||
EV230756 | 1/2 | Xylella fastidiosa | 9a5c |
(d)(U)C Filtration |
Mendes JS | 2016 | 56% | |
Study summaryFull title
All authors
Mendes JS, Santiago AS, Toledo MA, Horta MA, de Souza AA, Tasic L, de Souza AP
Journal
Front Microbiol
Abstract
The phytopathogen causes economic losses in important agricultural crops. Xylem vessel occlusion ca (show more...)
EV-METRIC
56% (90th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Cell culture supernatant
Sample origin
Control condition
Focus vesicles
outer membrane vesicles
Separation protocol
Separation protocol
(Differential) (ultra)centrifugation
Filtration Protein markers
EV: XF2491
non-EV: not specified Proteomics
yes
Show all info
Study aim
Identification of content (omics approaches)
Sample
Species
Xylella fastidiosa
Sample Type
Cell culture supernatant
EV-producing cells
9a5c
EV-harvesting Medium
Serum free medium
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 100,000 g and 150,000 g
Pelleting performed
Yes
Pelleting: time(min)
180
Pelleting: rotor type
L8-80 M
Pelleting: speed (g)
100000
Wash: volume per pellet (ml)
40
Wash: time (min)
120
Wash: speed (g)
100000
Filtration steps
0.2 or 0.22 μm
Characterization: Protein analysis
Protein Concentration Method
BCA
Western Blot
Detected EV-associated proteins
XF2491
Not detected EV-associated proteins
not specified
Detected contaminants
not specified
Not detected contaminants
not specified
Proteomics database
Unicamp
Characterization: Lipid analysis
No
Characterization: Particle analysis
EM
EM-type
Transmission-EM
Image type
Wide-field
|
||||||||
EV230045 | 1/4 | Homo sapiens | hMSC-TERT |
(d)(U)C Filtration |
L Ramos T | 2016 | 56% | |
Study summaryFull title
All authors
L Ramos T, Sánchez-Abarca LI, Muntión S, Preciado S, Puig N, López-Ruano G, Hernández-Hernández Á, Redondo A, Ortega R, Rodríguez C, Sánchez-Guijo F, del Cañizo C
Journal
Cell Commun Signal
Abstract
Human mesenchymal stromal cells (hMSC) are multipotent cells with both regenerative and immunomodula (show more...)
EV-METRIC
56% (90th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Cell culture supernatant
Sample origin
Control condition
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
(Differential) (ultra)centrifugation
Filtration Protein markers
EV: CD63/ CD81/ CD9/ CD73
non-EV: None Proteomics
no
Show all info
Study aim
Biomarker
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
EV-producing cells
hMSC-TERT
EV-harvesting Medium
Serum free medium
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 800 g and 10,000 g
Between 100,000 g and 150,000 g Pelleting performed
Yes
Pelleting: rotor type
70ti
Pelleting: speed (g)
100000
Wash: volume per pellet (ml)
not reported
Wash: time (min)
70
Wash: Rotor Type
70ti
Wash: speed (g)
100000
Filtration steps
0.2 or 0.22 µm
Characterization: Protein analysis
Protein Concentration Method
Bradford
Western Blot
Detected EV-associated proteins
CD63/ CD81/ CD73
Flow cytometry
Type of Flow cytometry
FACSCanto II (BD)
Calibration bead size
0.5/ 0.9/ 3
Detected EV-associated proteins
CD9/ CD81
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Mean
Reported size (nm)
159.7
EV concentration
Yes
EM
EM-type
Transmission-EM
Image type
Close-up, Wide-field
|
||||||||
EV230045 | 2/4 | Homo sapiens | hMSC-GFP |
(d)(U)C Filtration |
L Ramos T | 2016 | 56% | |
Study summaryFull title
All authors
L Ramos T, Sánchez-Abarca LI, Muntión S, Preciado S, Puig N, López-Ruano G, Hernández-Hernández Á, Redondo A, Ortega R, Rodríguez C, Sánchez-Guijo F, del Cañizo C
Journal
Cell Commun Signal
Abstract
Human mesenchymal stromal cells (hMSC) are multipotent cells with both regenerative and immunomodula (show more...)
EV-METRIC
56% (90th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Cell culture supernatant
Sample origin
Control condition
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
(Differential) (ultra)centrifugation
Filtration Protein markers
EV: CD63/ CD81/ CD73
non-EV: None Proteomics
no
Show all info
Study aim
Biomarker
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
EV-producing cells
hMSC-GFP
EV-harvesting Medium
Serum free medium
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 800 g and 10,000 g
Between 100,000 g and 150,000 g Pelleting performed
Yes
Pelleting: rotor type
70ti
Pelleting: speed (g)
100000
Wash: volume per pellet (ml)
not reported
Wash: time (min)
70
Wash: Rotor Type
70ti
Wash: speed (g)
100000
Filtration steps
0.2 or 0.22 µm
Characterization: Protein analysis
Protein Concentration Method
Bradford
Western Blot
Detected EV-associated proteins
CD63/ CD81/ CD73
Flow cytometry
Type of Flow cytometry
FACSCanto II (BD)
Calibration bead size
0.5/ 0.9/ 3
Detected EV-associated proteins
CD63/ CD81
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Mean
Reported size (nm)
136.6
EV concentration
Yes
EM
EM-type
Transmission-EM
Image type
Close-up, Wide-field
|
||||||||
EV230045 | 3/4 | Homo sapiens | HS-5 |
(d)(U)C Filtration |
L Ramos T | 2016 | 56% | |
Study summaryFull title
All authors
L Ramos T, Sánchez-Abarca LI, Muntión S, Preciado S, Puig N, López-Ruano G, Hernández-Hernández Á, Redondo A, Ortega R, Rodríguez C, Sánchez-Guijo F, del Cañizo C
Journal
Cell Commun Signal
Abstract
Human mesenchymal stromal cells (hMSC) are multipotent cells with both regenerative and immunomodula (show more...)
EV-METRIC
56% (90th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Cell culture supernatant
Sample origin
Control condition
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
(Differential) (ultra)centrifugation
Filtration Protein markers
EV: CD63/ CD81/ CD73
non-EV: None Proteomics
no
Show all info
Study aim
Biomarker
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
EV-producing cells
HS-5
EV-harvesting Medium
Serum free medium
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 800 g and 10,000 g
Between 100,000 g and 150,000 g Pelleting performed
Yes
Pelleting: rotor type
70ti
Pelleting: speed (g)
100000
Wash: volume per pellet (ml)
not reported
Wash: time (min)
70
Wash: Rotor Type
70ti
Wash: speed (g)
100000
Filtration steps
0.2 or 0.22 µm
Characterization: Protein analysis
Protein Concentration Method
Bradford
Western Blot
Detected EV-associated proteins
CD63/ CD81/ CD73
Flow cytometry
Type of Flow cytometry
FACSCanto II (BD)
Calibration bead size
0.5/ 0.9/ 3
Detected EV-associated proteins
CD63/ CD81
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Mean
Reported size (nm)
125.6
EV concentration
Yes
EM
EM-type
Transmission-EM
Image type
Close-up, Wide-field
|
||||||||
EV230045 | 4/4 | Homo sapiens | K562 |
(d)(U)C Filtration |
L Ramos T | 2016 | 56% | |
Study summaryFull title
All authors
L Ramos T, Sánchez-Abarca LI, Muntión S, Preciado S, Puig N, López-Ruano G, Hernández-Hernández Á, Redondo A, Ortega R, Rodríguez C, Sánchez-Guijo F, del Cañizo C
Journal
Cell Commun Signal
Abstract
Human mesenchymal stromal cells (hMSC) are multipotent cells with both regenerative and immunomodula (show more...)
EV-METRIC
56% (90th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Cell culture supernatant
Sample origin
Control condition
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
(Differential) (ultra)centrifugation
Filtration Protein markers
EV: CD63/ CD81/ CD73
non-EV: None Proteomics
no
Show all info
Study aim
Biomarker
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
EV-producing cells
K562
EV-harvesting Medium
Serum free medium
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 800 g and 10,000 g
Between 100,000 g and 150,000 g Pelleting performed
Yes
Pelleting: rotor type
70ti
Pelleting: speed (g)
100000
Wash: volume per pellet (ml)
not reported
Wash: time (min)
70
Wash: Rotor Type
70ti
Wash: speed (g)
100000
Filtration steps
0.2 or 0.22 µm
Characterization: Protein analysis
Protein Concentration Method
Bradford
Western Blot
Detected EV-associated proteins
CD63/ CD81/ CD73
Flow cytometry
Type of Flow cytometry
FACSCanto II (BD)
Calibration bead size
0.5/ 0.9/ 3
Detected EV-associated proteins
CD63/ CD81
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Mean
Reported size (nm)
122
EV concentration
Yes
EM
EM-type
Transmission-EM
Image type
Close-up, Wide-field
|
||||||||
EV210035 | 1/2 | Homo sapiens | A-431 |
(d)(U)C Filtration |
van Dommelen, Susan M | 2016 | 56% | |
Study summaryFull title
All authors
Susan M van Dommelen, Roy van der Meel, Wouter W van Solinge, Maria Coimbra, Pieter Vader, Raymond M Schiffelers
Journal
Nanomedicine
Abstract
Aim: Extracellular vesicles (EVs) are attractive candidates for biomarker research, because their co (show more...)
EV-METRIC
56% (90th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Cell culture supernatant
Sample origin
Control condition
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
(Differential) (ultra)centrifugation
Filtration Protein markers
EV: TSG101/ Alix/ Cetuximab/ EGFR/ pEGFR/ Akt/ pAkt/ -actin/ CD9
non-EV: Lamin-A/ Lamin-C/ ATP5-A/ Tom20 Proteomics
no
Show all info
Study aim
Function/Identification of content (omics approaches)
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
EV-producing cells
A-431
EV-harvesting Medium
Serum free medium
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Below or equal to 800 g
Between 800 g and 10,000 g Between 100,000 g and 150,000 g Pelleting performed
Yes
Pelleting: time(min)
70
Pelleting: rotor type
JA-30.50
Pelleting: speed (g)
100000
Wash: volume per pellet (ml)
Not specified
Wash: time (min)
70
Wash: Rotor Type
JA-30.50
Wash: speed (g)
100000
Filtration steps
0.22µm or 0.2µm
Characterization: Protein analysis
Protein Concentration Method
microBCA
Western Blot
Detected EV-associated proteins
CD9/ EGFR/ pEGFR/ Akt/ pAkt/ -actin/ TSG101/ Alix
Not detected EV-associated proteins
Cetuximab
Not detected contaminants
Lamin-A/ Lamin-C/ ATP5-A/ Tom20
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Not Reported
EV concentration
Yes
|
||||||||
EV160008 | 1/3 | Rattus norvegicus | Primary pancreatic islets | (d)(U)C | Cianciaruso C | 2016 | 55% | |
Study summaryFull title
All authors
Cianciaruso C, Phelps EA, Pasquier M, Hamelin R, Demurtas D, Alibashe Ahmed M, Piemonti L, Hirosue S, Swartz MA, De Palma M, Hubbell JA, Baekkeskov S
Journal
Diabetes
Abstract
The target autoantigens in several organ-specific autoimmune diseases, including type 1 diabetes (T1 (show more...)
EV-METRIC
55% (88th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Cell culture supernatant
Sample origin
Control condition
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
(d)(U)C
Protein markers
EV: GAD67/ GAD65/ Insulin/ PDI/ Flotillin-1/ CD9/ IA-2
non-EV: Gp96/ Calreticulin Proteomics
yes
Show all info
Study aim
Function, Biogenesis/cargo sorting, Identification of content (omics approaches)
Sample
Species
Rattus norvegicus
Sample Type
Cell culture supernatant
EV-producing cells
Primary pancreatic islets
EV-harvesting Medium
EV-depleted serum
Preparation of EDS
overnight (16h) at >=100,000g
Cell viability (%)
NA
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Below or equal to 800 g
Between 800 g and 10,000 g Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Pelleting performed
Yes
Pelleting: time(min)
70
Pelleting: speed (g)
110000
Wash: time (min)
70
Wash: speed (g)
110000
Characterization: Protein analysis
Protein Concentration Method
BCA
Western Blot
Detected EV-associated proteins
CD9, Flotillin-1, GAD65, GAD67, IA-2, PDI
Not detected contaminants
Calreticulin, Gp96
ELISA
Proteomics database
No
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Mode
Reported size (nm)
139
EM
EM-type
Transmission-EM
Image type
Wide-field
Report size (nm)
120
|
||||||||
EV160005 | 1/2 | Homo sapiens | Bronchoalveolar lavage fluid |
(d)(U)C Filtration |
Héliot A | 2016 | 55% | |
Study summaryFull title
All authors
Héliot A, Landkocz Y, Roy Saint-Georges F, Gosset P, Billet S, Shirali P, Courcot D, Martin PJ
Journal
Int J Hyg Environ Health
Abstract
Cigarette smoking is a habit that has spread all over the world and is a significant risk factor for (show more...)
EV-METRIC
55% (57th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Bronchoalveolar lavage fluid
Sample origin
Control condition
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
(d)(U)C
Filtration Adj. k-factor
126 (pelleting) / 126 (washing)
Protein markers
EV: CD81/ CD63/ CD9
non-EV: None Proteomics
no
Show all info
Study aim
Function, Biomarker
Sample
Species
Homo sapiens
Sample Type
Bronchoalveolar lavage fluid
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Below or equal to 800 g
Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Pelleting performed
Yes
Pelleting: time(min)
70
Pelleting: rotor type
SW 55 Ti
Pelleting: speed (g)
110000
Pelleting: adjusted k-factor
126.0
Wash: time (min)
70
Wash: Rotor Type
SW 55 Ti
Wash: speed (g)
110000
Wash: adjusted k-factor
126.0
Filtration steps
0.22µm or 0.2µm
Characterization: Protein analysis
Protein Concentration Method
Not determined
Western Blot
Detected EV-associated proteins
CD9, CD63, CD81
Characterization: RNA analysis
Database
No
Proteinase treatment
No
RNAse treatment
No
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Size range/distribution;Mean
Reported size (nm)
138.1±2.2
EV concentration
Yes
Particle yield
2.48E+11 particles/ml start sample
EM
EM-type
Transmission-EM
Image type
Close-up, Wide-field
Report size (nm)
30-200
Extra information
UC rotors added to the EV-TRACK entry after publication
|
||||||||
EV160005 | 2/2 | Homo sapiens | Bronchoalveolar lavage fluid |
(d)(U)C Filtration |
Héliot A | 2016 | 55% | |
Study summaryFull title
All authors
Héliot A, Landkocz Y, Roy Saint-Georges F, Gosset P, Billet S, Shirali P, Courcot D, Martin PJ
Journal
Int J Hyg Environ Health
Abstract
Cigarette smoking is a habit that has spread all over the world and is a significant risk factor for (show more...)
EV-METRIC
55% (57th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Bronchoalveolar lavage fluid
Sample origin
Smokers
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
(d)(U)C
Filtration Adj. k-factor
126 (pelleting) / 126 (washing)
Protein markers
EV: CD81/ CD63/ CD9
non-EV: None Proteomics
no
Show all info
Study aim
Function, Biomarker
Sample
Species
Homo sapiens
Sample Type
Bronchoalveolar lavage fluid
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Below or equal to 800 g
Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Pelleting performed
Yes
Pelleting: time(min)
70
Pelleting: rotor type
SW 55 Ti
Pelleting: speed (g)
110000
Pelleting: adjusted k-factor
126.0
Wash: time (min)
70
Wash: Rotor Type
SW 55 Ti
Wash: speed (g)
110000
Wash: adjusted k-factor
126.0
Filtration steps
0.22µm or 0.2µm
Characterization: Protein analysis
Protein Concentration Method
Not determined
Western Blot
Detected EV-associated proteins
CD9, CD63, CD81
Characterization: RNA analysis
Database
No
Proteinase treatment
No
RNAse treatment
No
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Size range/distribution;Mean
Reported size (nm)
139.8±3.3
EV concentration
Yes
Particle yield
1.94E+11 particles/ml start sample
EM
EM-type
Transmission-EM
Image type
Close-up, Wide-field
Report size (nm)
30-200
Extra information
UC rotors added to the EV-TRACK entry after publication
|
||||||||
EV160004 | 1/5 | Equus caballus | Synovial fluid |
DG (d)(U)C |
Boere J | 2016 | 55% | |
Study summaryFull title
All authors
Boere J, van de Lest CH, Libregts SF, Arkesteijn GJ, Geerts WJ, Nolte-'t Hoen EN, Malda J, van Weeren PR, Wauben MH
Journal
J Extracell Vesicles
Abstract
Extracellular vesicles (EVs) in synovial fluid (SF) are gaining increased recognition as important f (show more...)
EV-METRIC
55% (71st percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Synovial fluid
Sample origin
Control condition
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
DG
(d)(U)C Adj. k-factor
83.68 (pelleting)
Protein markers
EV: CD44/ CD9
non-EV: None Proteomics
no
Show all info
Study aim
Biomarker, New methodological development
Sample
Species
Equus caballus
Sample Type
Synovial fluid
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 800 g and 10,000 g
Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Equal to or above 150,000 g Pelleting performed
Yes
Pelleting: time(min)
65
Pelleting: rotor type
SW 60 Ti
Pelleting: speed (g)
200000
Pelleting: adjusted k-factor
83.68
Density gradient
Type
Continuous
Number of initial discontinuous layers
15
Highest density fraction
0.51
Sample volume (mL)
1.75
Orientation
Bottom-up (sample migrates upwards)
Rotor type
SW 40 Ti
Speed (g)
200000
Duration (min)
960
Fraction volume (mL)
1
Fraction processing
Centrifugation
Pelleting: volume per fraction
19
Pelleting: duration (min)
60
Pelleting: rotor type
SW 28
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
253.9
Characterization: Protein analysis
Protein Concentration Method
Not determined
Western Blot
Detected EV-associated proteins
CD9, CD44
Flow cytometry
Type of Flow cytometry
BD-Influx
Hardware adaptation to ~100nm EV's
optimized jet-in-air-based BD Influx flow cytometer
Calibration bead size
0.1,0.2
Characterization: Lipid analysis
No
Characterization: Particle analysis
Particle analysis: flow cytometry
Flow cytometer type
BD-Influx
Hardware adjustment
optimized jet-in-air-based BD Influx flow cytometer
Calibration bead size
0.1;0.2
EV concentration
Yes
Particle yield
7.00E+08 particles/ml start sample
Extra information
Importantly, synovial fluid samples were pre-treated with hyaluronidase prior to ultracentrifugation. Concentration calculated as sum of EVs recovered after all pelleting steps (10K, 100K, 200K).
|
||||||||
EV160004 | 3/5 | Equus caballus | Synovial fluid |
DG (d)(U)C |
Boere J | 2016 | 55% | |
Study summaryFull title
All authors
Boere J, van de Lest CH, Libregts SF, Arkesteijn GJ, Geerts WJ, Nolte-'t Hoen EN, Malda J, van Weeren PR, Wauben MH
Journal
J Extracell Vesicles
Abstract
Extracellular vesicles (EVs) in synovial fluid (SF) are gaining increased recognition as important f (show more...)
EV-METRIC
55% (71st percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Synovial fluid
Sample origin
Control condition
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
DG
(d)(U)C Adj. k-factor
167.3 (pelleting)
Protein markers
EV: CD44/ CD9
non-EV: None Proteomics
no
Show all info
Study aim
Biomarker, New methodological development
Sample
Species
Equus caballus
Sample Type
Synovial fluid
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 800 g and 10,000 g
Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Pelleting performed
Yes
Pelleting: time(min)
65
Pelleting: rotor type
SW 60 Ti
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
167.3
Density gradient
Type
Continuous
Number of initial discontinuous layers
15
Highest density fraction
0.51
Sample volume (mL)
1.75
Orientation
Bottom-up (sample migrates upwards)
Rotor type
SW 40 Ti
Speed (g)
200000
Duration (min)
960
Fraction volume (mL)
1
Fraction processing
Centrifugation
Pelleting: volume per fraction
19
Pelleting: duration (min)
60
Pelleting: rotor type
SW 28
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
253.9
Characterization: Protein analysis
Protein Concentration Method
Not determined
Western Blot
Detected EV-associated proteins
CD9, CD44
Flow cytometry
Type of Flow cytometry
BD-Influx
Hardware adaptation to ~100nm EV's
optimized jet-in-air-based BD Influx flow cytometer
Calibration bead size
0.1,0.2
Characterization: Lipid analysis
No
Characterization: Particle analysis
Particle analysis: flow cytometry
Flow cytometer type
BD-Influx
Hardware adjustment
optimized jet-in-air-based BD Influx flow cytometer
Calibration bead size
0.1;0.2
EV concentration
Yes
Particle yield
7.00E+08 particles/ml start sample
Extra information
Importantly, synovial fluid samples were pre-treated with hyaluronidase prior to ultracentrifugation. Concentration calculated as sum of EVs recovered after all pelleting steps (10K, 100K, 200K).
|
||||||||
EV160004 | 4/5 | Equus caballus | Synovial fluid |
DG (d)(U)C |
Boere J | 2016 | 55% | |
Study summaryFull title
All authors
Boere J, van de Lest CH, Libregts SF, Arkesteijn GJ, Geerts WJ, Nolte-'t Hoen EN, Malda J, van Weeren PR, Wauben MH
Journal
J Extracell Vesicles
Abstract
Extracellular vesicles (EVs) in synovial fluid (SF) are gaining increased recognition as important f (show more...)
EV-METRIC
55% (71st percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Synovial fluid
Sample origin
Control condition
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
DG
(d)(U)C Adj. k-factor
1673 (pelleting)
Protein markers
EV: CD44/ CD9
non-EV: None Proteomics
no
Show all info
Study aim
Biomarker, New methodological development
Sample
Species
Equus caballus
Sample Type
Synovial fluid
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 800 g and 10,000 g
Between 10,000 g and 50,000 g Pelleting performed
Yes
Pelleting: time(min)
35
Pelleting: rotor type
SW 60 Ti
Pelleting: speed (g)
10000
Pelleting: adjusted k-factor
1673.
Density gradient
Type
Continuous
Number of initial discontinuous layers
15
Highest density fraction
0.51
Sample volume (mL)
1.75
Orientation
Bottom-up (sample migrates upwards)
Rotor type
SW 40 Ti
Speed (g)
200000
Duration (min)
960
Fraction volume (mL)
1
Fraction processing
Centrifugation
Pelleting: volume per fraction
19
Pelleting: duration (min)
60
Pelleting: rotor type
SW 28
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
253.9
Characterization: Protein analysis
Protein Concentration Method
Not determined
Western Blot
Detected EV-associated proteins
CD9, CD44
Flow cytometry
Type of Flow cytometry
BD-Influx
Hardware adaptation to ~100nm EV's
optimized jet-in-air-based BD Influx flow cytometer
Calibration bead size
0.1,0.2
Characterization: Lipid analysis
No
Characterization: Particle analysis
Particle analysis: flow cytometry
Flow cytometer type
BD-Influx
Hardware adjustment
optimized jet-in-air-based BD Influx flow cytometer
Calibration bead size
0.1;0.2
EV concentration
Yes
Particle yield
7.00E+08 particles/ml start sample
Extra information
Importantly, synovial fluid samples were pre-treated with hyaluronidase prior to ultracentrifugation. Concentration calculated as sum of EVs recovered after all pelleting steps (10K, 100K, 200K).
|
||||||||
EV210488 | 1/11 | Homo sapiens | Blood plasma |
(d)(U)C SEC (non-commercial) DG |
van Eijndhoven MA | 2016 | 50% | |
Study summaryFull title
All authors
van Eijndhoven MA, Zijlstra JM, Groenewegen NJ, Drees EE, van Niele S, Baglio SR, Koppers-Lalic D, van der Voorn H, Libregts SF, Wauben MH, de Menezes RX, van Weering JR, Nieuwland R, Visser L, van den Berg A, de Jong D, Pegtel DM
Journal
JCI insight
Abstract
Cell-free circulating nucleic acids, including 22-nt microRNAs (miRNAs), represent noninvasive bioma (show more...)
EV-METRIC
50% (82nd percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Blood plasma
Sample origin
Control condition
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
(Differential) (ultra)centrifugation
Size-exclusion chromatography (non-commercial) Density gradient Protein markers
EV: None
non-EV: None Proteomics
no
Show all info
Study aim
Biomarker
Sample
Species
Homo sapiens
Sample Type
Blood plasma
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Below or equal to 800 g
Between 800 g and 10,000 g Pelleting performed
No
Density gradient
Only used for validation of main results
Yes
Type
Discontinuous
Number of initial discontinuous layers
15
Lowest density fraction
0.4M
Highest density fraction
2M
Orientation
Bottom-up
Rotor type
SW 40 Ti
Speed (g)
192000
Duration (min)
840-1200
Fraction volume (mL)
1
Fraction processing
None
Size-exclusion chromatography
Used for validation?
Yes
Total column volume (mL)
10
Sample volume/column (mL)
1.5
Characterization: Protein analysis
None
Protein Concentration Method
Not determined
Characterization: RNA analysis
RNA analysis
Type
(RT)(q)PCR/ RNA sequencing/ Capillary electrophoresis (e.g. Bioanalyzer)
Database
No
Proteinase treatment
No
RNAse treatment
No
Characterization: Lipid analysis
No
Characterization: Particle analysis
TRPS
Report type
Size range/distribution
Reported size (nm)
50-500
EV concentration
Yes
Particle analysis: flow cytometry
Flow cytometer type
Influx (Becton Dickinson)
Hardware adjustment
BD Influx cell sorter Influx cell sorter , which was equipped by Cytopeia and BD with: 488nm laser (200 mW)/ 561-nm laser (100 mW)/ 635-nm CUBE laser (30 mW) / SPD, containing a Mitutoyo Plan Apo 20 lens (NA 0.42), a 0.7-mm pinhole and a polarization unit with two FSC PMTs/ Optical filters/ amplifier board with 2 'fast' preamplifiers (0.12 s) and 6 'slow' preamplifiers (1.2 s)/ and Spigot software, version 6.1.
Calibration bead size
0.1
Report type
Not Reported
EV concentration
Yes
EM
EM-type
Transmission-EM
Image type
Wide-field
|
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EV210101 | 3/6 | Homo sapiens | Blood plasma |
(d)(U)C SEC (non-commercial) Filtration |
Welton, Joanne Louise | 2016 | 50% | |
Study summaryFull title
All authors
Joanne Louise Welton, Paul Brennan, Mark Gurney, Jason Paul Webber, Lisa Kate Spary, David Gil Carton, Juan Manuel Falcón-Pérez, Sean Peter Walton, Malcolm David Mason, Zsuzsanna Tabi, Aled Clayton
Journal
J Extracell Vesicles
Abstract
Proteomics analysis of biofluid-derived vesicles holds enormous potential for discovering non-invasi (show more...)
EV-METRIC
50% (82nd percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Blood plasma
Sample origin
Control condition
Focus vesicles
exosome
Separation protocol
Separation protocol
(d)(U)C
Size-exclusion chromatography (non-commercial) Filtration Protein markers
EV: CD81/ CD9
non-EV: Albumin/ ApoB Proteomics
no
Show all info
Study aim
Identification of content (omics approaches)/Technical analysis comparing/optimizing EV-related methods
Sample
Species
Homo sapiens
Sample Type
Blood plasma
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Below or equal to 800 g
Between 800 g and 10,000 g Equal to or above 150,000 g Pelleting performed
Yes
Pelleting: time(min)
120
Pelleting: rotor type
TLA-110
Pelleting: speed (g)
200000
Filtration steps
0.22µm or 0.2µm
Size-exclusion chromatography
Total column volume (mL)
12
Sample volume/column (mL)
1.5
Resin type
Sepharose CL-2B
Characterization: Protein analysis
Protein Concentration Method
Other;Spectrophotometry
ELISA
Detected EV-associated proteins
CD81/ CD9
Detected contaminants
ApoB
Detected EV-associated proteins
SOMAscan multiplex assay
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Mean
Reported size (nm)
86.47
EV concentration
Yes
Particle yield
As the number of particles per µg protein;Yes, other: 5000000000
EM
EM-type
Cryo-EM
Image type
Close-up
|
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EV160007 | 1/3 | Homo sapiens | Blood plasma |
(d)(U)C Filtration SEC |
Hong CS | 2016 | 50% | |
Study summaryFull title
All authors
Hong CS, Funk S, Muller L, Boyiadzis M, Whiteside TL
Journal
J Extracell Vesicles
Abstract
OBJECTIVE:
Isolation from human plasma of exosomes that retain functional and morphological integri (show more...)
EV-METRIC
50% (82nd percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Blood plasma
Sample origin
Acute myeloid leukemia
Focus vesicles
exosome
Separation protocol
Separation protocol
(d)(U)C
Filtration SEC Protein markers
EV: TSG101/ PD-1/ CD123/ CD96/ CD44/ CD34/ Pro-TGFbeta1 LAP/ Pro-TGFbeta1LAP/ PD-L1/ CLL-1/ CD9
non-EV: None Proteomics
no
Show all info
Study aim
Function, Biomarker, New methodological development, Identification of content (omics approaches)
Sample
Species
Homo sapiens
Sample Type
Blood plasma
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 800 g and 10,000 g
Between 10,000 g and 50,000 g Pelleting performed
No
Filtration steps
0.22µm or 0.2µm
Size-exclusion chromatography
Total column volume (mL)
10
Sample volume/column (mL)
1
Resin type
Sepharose CL-2B
Characterization: Protein analysis
Protein Concentration Method
BCA
Protein Yield (µg)
66
Western Blot
Detected EV-associated proteins
CD9, TSG101, CD44, CD34, CD123, CD96, CLL-1, Pro-TGFbeta1 LAP, PD-1, PD-L1
Characterization: Lipid analysis
No
Characterization: Particle analysis
TRPS
Report type
Mean
Reported size (nm)
77-92
EV concentration
Yes
Particle yield
8.90E+10 particles/ml start sample
EM
EM-type
Transmission-EM
Image type
Wide-field
|
||||||||
EV160007 | 2/3 | Homo sapiens | Blood plasma |
(d)(U)C Filtration SEC |
Hong CS | 2016 | 50% | |
Study summaryFull title
All authors
Hong CS, Funk S, Muller L, Boyiadzis M, Whiteside TL
Journal
J Extracell Vesicles
Abstract
OBJECTIVE:
Isolation from human plasma of exosomes that retain functional and morphological integri (show more...)
EV-METRIC
50% (82nd percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Blood plasma
Sample origin
Head and neck squamous cell carcinoma
Focus vesicles
exosome
Separation protocol
|