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You searched for: EV190006 (EV-TRACK ID)
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Details | EV-TRACK ID | Experiment nr. | Species | Sample type | separation protocol | First author | Year | EV-METRIC |
---|---|---|---|---|---|---|---|---|
EV190006 | 1/2 | Homo sapiens | Cell culture supernatant |
DG (d)(U)C |
Altei, Wanessa F. | 2020 | 100% | |
Study summaryFull title
All authors
Wanessa F Altei, Bianca C Pachane, Patty K Dos Santos, Lígia N M Ribeiro, Bong Hwan Sung, Alissa M Weaver, Heloisa S Selistre-de-Araújo
Journal
Cell Commun Signal
Abstract
Background: Extracellular vesicles (EVs) are lipid-bound particles that are naturally released from (show more...)
EV-METRIC
100% (99th 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
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
Cell Name
MDAMB231
Sample origin
immortalized
Focus vesicles
exosome
Separation protocol
Separation protocol
Density gradient
(Differential) (ultra)centrifugation Protein markers
EV: Flotillin1/ CD63/ Alix/ integrin-alpha5/ integrin-alpha2/ integrin-alphaV/ integrin-beta1/ integrin-beta3/ FN1/ COL1
non-EV: Calnexin Proteomics
no
EV density (g/ml)
1.11
Show all info
Study aim
Biogenesis/cargo sorting/Mechanism of uptake/transfer
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
Sample Condition
immortalized
EV-producing cells
MDAMB231
EV-harvesting Medium
Serum free medium
Cell viability
Yes
Cell viability (%)
Yes
Separation Method
Differential ultracentrifugation
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 Obtain an EV pellet :
Yes
Pelleting: time(min)
4
Pelleting: rotor type
SW 32 Ti
Pelleting: speed (g)
100000
Density gradient
Density medium
Iodixanol
Type
Discontinuous
Number of initial discontinuous layers
4
Lowest density fraction
5%
Highest density fraction
40%
Total gradient volume, incl. sample (mL)
12
Sample volume (mL)
3
Orientation
Bottom-up
Rotor type
Type 40
Speed (g)
100000
Duration (min)
18
Fraction volume (mL)
1
Fraction processing
Centrifugation
Pelleting: volume per fraction
1
Pelleting: duration (min)
180
Pelleting: rotor type
TLA-100.3
Pelleting: speed (g)
100000
Pelleting-wash: volume per pellet (mL)
3
Pelleting-wash: duration (min)
180
Pelleting-wash: speed (g)
TLA-100.3
Density cushion
Density medium
EV-subtype
Distinction between multiple subtypes
Used subtypes
Characterization: Protein analysis
Protein Concentration Method
microBCA
Western Blot
Detected EV-associated proteins
Flotillin1/ CD63/ Alix/ integrin-alpha5/ integrin-alpha2/ integrin-alphaV/ integrin-beta1/ integrin-beta3/ FN1/ COL1
Not detected EV-associated proteins
Not detected contaminants
Calnexin
Flow cytometry
Hardware adjustments
Characterization: Particle analysis
NTA
Report type
Median
Reported size (nm)
107
EV concentration
Yes
Particle yield
Yes, as number of particles per million cells 5.50E+08
EM
EM-type
Transmission-EM
Image type
Close-up, Wide-field
Report size (nm)
|
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EV190006 | 2/2 | Homo sapiens | Cell culture supernatant | (d)(U)C | Altei, Wanessa F. | 2020 | 78% | |
Study summaryFull title
All authors
Wanessa F Altei, Bianca C Pachane, Patty K Dos Santos, Lígia N M Ribeiro, Bong Hwan Sung, Alissa M Weaver, Heloisa S Selistre-de-Araújo
Journal
Cell Commun Signal
Abstract
Background: Extracellular vesicles (EVs) are lipid-bound particles that are naturally released from (show more...)
EV-METRIC
78% (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
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
Cell Name
MDAMB231
Sample origin
immortalized
Focus vesicles
(shedding) microvesicle
Separation protocol
Separation protocol
(Differential) (ultra)centrifugation
Protein markers
EV: Flotillin1/ CD63/ Alix/ integrin-alpha5/ integrin-alpha2/ integrin-alphaV/ integrin-beta1/ integrin-beta3/ FN1/ COL1
non-EV: Calnexin Proteomics
no
EV density (g/ml)
Show all info
Study aim
Biogenesis/cargo sorting/Mechanism of uptake/transfer
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
Sample Condition
immortalized
EV-producing cells
MDAMB231
EV-harvesting Medium
Serum free medium
Cell viability
Yes
Cell viability (%)
Yes
Separation Method
Differential ultracentrifugation
centrifugation steps
Below or equal to 800 g
Between 800 g and 10,000 g Between 10,000 g and 50,000 g Obtain an EV pellet :
Yes
Pelleting: time(min)
4
Pelleting: rotor type
SW 32 Ti
Pelleting: speed (g)
10000
Density gradient
Only used for validation of main results
Density medium
Type
Number of initial discontinuous layers
Lowest density fraction
Highest density fraction
Total gradient volume, incl. sample (mL)
Sample volume (mL)
Orientation
Rotor type
Speed (g)
Duration (min)
Fraction volume (mL)
Fraction processing
Pelleting: volume per fraction
Pelleting: duration (min)
Pelleting: rotor type
Pelleting: speed (g)
Pelleting-wash: volume per pellet (mL)
Pelleting-wash: duration (min)
Pelleting-wash: speed (g)
Density cushion
Density medium
EV-subtype
Distinction between multiple subtypes
Used subtypes
Characterization: Protein analysis
Protein Concentration Method
microBCA
Western Blot
Detected EV-associated proteins
Flotillin1/ integrin-alpha5/ integrin-beta1
Not detected EV-associated proteins
CD63/ Alix/ FN1/ COL1
Not detected contaminants
Calnexin
Flow cytometry
Hardware adjustments
Characterization: Particle analysis
NTA
Report type
Median
Reported size (nm)
206
EV concentration
Yes
Particle yield
Yes, as number of particles per million cells 9.00E+08
EM
EM-type
Transmission-EM
Image type
Close-up, Wide-field
Report size (nm)
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