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You searched for: EV200093 (EV-TRACK ID)
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Showing 1 - 10 of 10
Details | EV-TRACK ID | Experiment nr. | Species | Sample type | Separation protocol | First author | Year | EV-METRIC |
---|---|---|---|---|---|---|---|---|
EV200093 | 10/10 | Homo sapiens | Blood plasma |
(d)(U)C Filtration DG |
Dong, Liang | 2021 | 63% | |
Study summaryFull title
All authors
Liang Dong, Richard C. Zieren, Kengo Horie, Chi‐Ju Kim, Emily Mallick, Yuezhou Jing, Mingxiao Feng, Morgan D. Kuczler, Jordan Green, Sarah R. Amend, Kenneth W. Witwer, Theo M. de Reijke, Yoon‐Kyoung Cho, Kenneth J. Pienta, Wei Xue
Journal
J Extracell Vesicles
Abstract
One of the challenges that restricts the evolving extracellular vesicle (EV) research field is the l (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
Control condition
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
(d)(U)C
Filtration DG Protein markers
EV: CD81/ Flotillin1
non-EV: ApoA1 Proteomics
no
EV density (g/ml)
1.10-1.15
Show all info
Study aim
Technical analysis comparing/optimizing EV-related methods
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
Density gradient
Only used for validation of main results
Yes
Type
Continuous
Lowest density fraction
5%
Highest density fraction
60%
Total gradient volume, incl. sample (mL)
38
Sample volume (mL)
2
Orientation
Bottom-up
Rotor type
SW 32 Ti
Speed (g)
100000
Duration (min)
230
Fraction volume (mL)
4.75
Fraction processing
Centrifugation
Pelleting: volume per fraction
28
Pelleting: duration (min)
60
Pelleting: rotor type
Type 70 Ti
Pelleting: speed (g)
120000
Filtration steps
0.45µm > x > 0.22µm,
Characterization: Protein analysis
Protein Concentration Method
Not determined
Western Blot
Antibody details provided?
Yes
Antibody dilution provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
Flotillin1/ CD81
Not detected contaminants
ApoA1
Flow cytometry
Type of Flow cytometry
NanoFCM
Hardware adaptation to ~100nm EV's
Please refer to the publication below: Zhu S, Ma L, Wang S, et al. Light-scattering detection below the level of single fluorescent molecules for high-resolution characterization of functional nanopar
Calibration bead size
0.2
Antibody details provided?
No
Characterization: Lipid analysis
No
Characterization: Particle analysis
Particle analysis: flow cytometry
Flow cytometer type
NanoFCM
Hardware adjustment
Please refer to the publication below: Zhu S, Ma L, Wang S, et al. Light-scattering detection below the level of single fluorescent molecules for high-resolution characterization of functional nanoparticles. ACS nano. 2014 Oct 28;8(10):10998-1006.
Calibration bead size
0.2
Report type
Not Reported
|
||||||||
EV200093 | 1/10 | Homo sapiens | PC3 |
(d)(U)C Filtration |
Dong, Liang | 2021 | 56% | |
Study summaryFull title
All authors
Liang Dong, Richard C. Zieren, Kengo Horie, Chi‐Ju Kim, Emily Mallick, Yuezhou Jing, Mingxiao Feng, Morgan D. Kuczler, Jordan Green, Sarah R. Amend, Kenneth W. Witwer, Theo M. de Reijke, Yoon‐Kyoung Cho, Kenneth J. Pienta, Wei Xue
Journal
J Extracell Vesicles
Abstract
One of the challenges that restricts the evolving extracellular vesicle (EV) research field is the l (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
(d)(U)C
Filtration Protein markers
EV: CD81/ Flotillin1/ CD63/ CD9
non-EV: Calreticulin Proteomics
no
Show all info
Study aim
Technical analysis comparing/optimizing EV-related methods
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
EV-producing cells
PC3
EV-harvesting Medium
EV-depleted medium
Preparation of EDS
Commercial EDS
Cell viability (%)
99
Cell count
7.50E+08
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
Type 70 Ti
Pelleting: speed (g)
120000
Wash: volume per pellet (ml)
28
Wash: time (min)
120
Wash: Rotor Type
Type 70 Ti
Wash: speed (g)
120000
Filtration steps
0.45µm > x > 0.22µm,
Characterization: Protein analysis
Protein Concentration Method
microBCA
Western Blot
Antibody details provided?
Yes
Antibody dilution provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
Flotillin1/ CD63/ CD81
Not detected contaminants
Calreticulin
Flow cytometry
Type of Flow cytometry
NanoFCM
Hardware adaptation to ~100nm EV's
Please refer to the publication below: Zhu S, Ma L, Wang S, et al. Light-scattering detection below the level of single fluorescent molecules for high-resolution characterization of functional nanopar
Calibration bead size
0.2
Antibody details provided?
No
Detected EV-associated proteins
CD63/ CD9/ CD81
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Modus
Reported size (nm)
113.5
EV concentration
Yes
Particle yield
Yes, as number of particles per milliliter of starting sample 9.05E+07
Particle analysis: flow cytometry
Flow cytometer type
NanoFCM
Hardware adjustment
Please refer to the publication below: Zhu S, Ma L, Wang S, et al. Light-scattering detection below the level of single fluorescent molecules for high-resolution characterization of functional nanoparticles. ACS nano. 2014 Oct 28;8(10):10998-1006.
Calibration bead size
0.2
EV concentration
Yes
|
||||||||
EV200093 | 6/10 | Homo sapiens | Blood plasma |
(d)(U)C Filtration |
Dong, Liang | 2021 | 56% | |
Study summaryFull title
All authors
Liang Dong, Richard C. Zieren, Kengo Horie, Chi‐Ju Kim, Emily Mallick, Yuezhou Jing, Mingxiao Feng, Morgan D. Kuczler, Jordan Green, Sarah R. Amend, Kenneth W. Witwer, Theo M. de Reijke, Yoon‐Kyoung Cho, Kenneth J. Pienta, Wei Xue
Journal
J Extracell Vesicles
Abstract
One of the challenges that restricts the evolving extracellular vesicle (EV) research field is the l (show more...)
EV-METRIC
56% (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
Blood plasma
Sample origin
Control condition
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
(d)(U)C
Filtration Protein markers
EV: CD81/ Flotillin1/ CD63/ CD9
non-EV: ApoA1/ Calreticulin Proteomics
no
Show all info
Study aim
Technical analysis comparing/optimizing EV-related methods
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
Type 70 Ti
Pelleting: speed (g)
120000
Wash: volume per pellet (ml)
28
Wash: time (min)
120
Wash: Rotor Type
Type 70 Ti
Wash: speed (g)
120000
Filtration steps
0.45µm > x > 0.22µm,
Characterization: Protein analysis
Protein Concentration Method
BCA
Western Blot
Antibody details provided?
Yes
Antibody dilution provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
Flotillin1/ CD81
Detected contaminants
ApoA1
Not detected contaminants
Calreticulin
Flow cytometry
Type of Flow cytometry
NanoFCM
Hardware adaptation to ~100nm EV's
Please refer to the publication below: Zhu S, Ma L, Wang S, et al. Light-scattering detection below the level of single fluorescent molecules for high-resolution characterization of functional nanopar
Calibration bead size
0.2
Antibody details provided?
No
Detected EV-associated proteins
CD63/ CD9/ CD81
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Modus
Reported size (nm)
121.5
EV concentration
Yes
Particle yield
Yes, as number of particles per milliliter of starting sample 2.42E+09
Particle analysis: flow cytometry
Flow cytometer type
NanoFCM
Hardware adjustment
Please refer to the publication below: Zhu S, Ma L, Wang S, et al. Light-scattering detection below the level of single fluorescent molecules for high-resolution characterization of functional nanoparticles. ACS nano. 2014 Oct 28;8(10):10998-1006.
Calibration bead size
0.2
Report type
Modus
Reported size (nm)
58.25
EV concentration
Yes
|
||||||||
EV200093 | 2/10 | Homo sapiens | PC3 |
(d)(U)C Filtration Total Exosome Isolation |
Dong, Liang | 2021 | 50% | |
Study summaryFull title
All authors
Liang Dong, Richard C. Zieren, Kengo Horie, Chi‐Ju Kim, Emily Mallick, Yuezhou Jing, Mingxiao Feng, Morgan D. Kuczler, Jordan Green, Sarah R. Amend, Kenneth W. Witwer, Theo M. de Reijke, Yoon‐Kyoung Cho, Kenneth J. Pienta, Wei Xue
Journal
J Extracell Vesicles
Abstract
One of the challenges that restricts the evolving extracellular vesicle (EV) research field is the l (show more...)
EV-METRIC
50% (87th 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
Filtration Commercial method Protein markers
EV: CD81/ Flotillin1/ CD63/ CD9
non-EV: Calreticulin Proteomics
no
Show all info
Study aim
Technical analysis comparing/optimizing EV-related methods
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
EV-producing cells
PC3
EV-harvesting Medium
EV-depleted medium
Preparation of EDS
Commercial EDS
Cell viability (%)
99
Cell count
7.50E+08
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
0.45µm > x > 0.22µm,
Commercial kit
Total Exosome Isolation
Characterization: Protein analysis
Protein Concentration Method
microBCA
Western Blot
Antibody details provided?
Yes
Antibody dilution provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
Flotillin1/ CD63/ CD81
Not detected contaminants
Calreticulin
Flow cytometry
Type of Flow cytometry
NanoFCM
Hardware adaptation to ~100nm EV's
Please refer to the publication below: Zhu S, Ma L, Wang S, et al. Light-scattering detection below the level of single fluorescent molecules for high-resolution characterization of functional nanopar
Calibration bead size
0.2
Antibody details provided?
No
Detected EV-associated proteins
CD63/ CD9/ CD81
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Modus
Reported size (nm)
110.5
EV concentration
Yes
Particle yield
Yes, as number of particles per milliliter of starting sample 4.42E+08
Particle analysis: flow cytometry
Flow cytometer type
NanoFCM
Hardware adjustment
Please refer to the publication below: Zhu S, Ma L, Wang S, et al. Light-scattering detection below the level of single fluorescent molecules for high-resolution characterization of functional nanoparticles. ACS nano. 2014 Oct 28;8(10):10998-1006.
Calibration bead size
0.2
Report type
Modus
Reported size (nm)
59.75
EV concentration
Yes
|
||||||||
EV200093 | 3/10 | Homo sapiens | PC3 |
(d)(U)C Filtration Exodisc |
Dong, Liang | 2021 | 50% | |
Study summaryFull title
All authors
Liang Dong, Richard C. Zieren, Kengo Horie, Chi‐Ju Kim, Emily Mallick, Yuezhou Jing, Mingxiao Feng, Morgan D. Kuczler, Jordan Green, Sarah R. Amend, Kenneth W. Witwer, Theo M. de Reijke, Yoon‐Kyoung Cho, Kenneth J. Pienta, Wei Xue
Journal
J Extracell Vesicles
Abstract
One of the challenges that restricts the evolving extracellular vesicle (EV) research field is the l (show more...)
EV-METRIC
50% (87th 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
Filtration Exodisc Protein markers
EV: CD81/ Flotillin1/ CD63/ CD9
non-EV: Calreticulin Proteomics
no
Show all info
Study aim
Technical analysis comparing/optimizing EV-related methods
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
EV-producing cells
PC3
EV-harvesting Medium
EV-depleted medium
Preparation of EDS
Commercial EDS
Cell viability (%)
99
Cell count
7.50E+08
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
0.45µm > x > 0.22µm,
Other
Name other separation method
Exodisc
Characterization: Protein analysis
Protein Concentration Method
microBCA
Western Blot
Antibody details provided?
Yes
Antibody dilution provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
Flotillin1/ CD63/ CD81
Not detected contaminants
Calreticulin
Flow cytometry
Type of Flow cytometry
NanoFCM
Hardware adaptation to ~100nm EV's
Please refer to the publication below: Zhu S, Ma L, Wang S, et al. Light-scattering detection below the level of single fluorescent molecules for high-resolution characterization of functional nanopar
Calibration bead size
0.2
Antibody details provided?
No
Detected EV-associated proteins
CD63/ CD9/ CD81
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Modus
Reported size (nm)
115.5
EV concentration
Yes
Particle yield
Yes, as number of particles per milliliter of starting sample 2.19E+08
Particle analysis: flow cytometry
Flow cytometer type
NanoFCM
Hardware adjustment
Please refer to the publication below: Zhu S, Ma L, Wang S, et al. Light-scattering detection below the level of single fluorescent molecules for high-resolution characterization of functional nanoparticles. ACS nano. 2014 Oct 28;8(10):10998-1006.
Calibration bead size
0.2
Report type
Modus
Reported size (nm)
57.25
EV concentration
Yes
|
||||||||
EV200093 | 4/10 | Homo sapiens | PC3 |
(d)(U)C Filtration SEC and UF |
Dong, Liang | 2021 | 50% | |
Study summaryFull title
All authors
Liang Dong, Richard C. Zieren, Kengo Horie, Chi‐Ju Kim, Emily Mallick, Yuezhou Jing, Mingxiao Feng, Morgan D. Kuczler, Jordan Green, Sarah R. Amend, Kenneth W. Witwer, Theo M. de Reijke, Yoon‐Kyoung Cho, Kenneth J. Pienta, Wei Xue
Journal
J Extracell Vesicles
Abstract
One of the challenges that restricts the evolving extracellular vesicle (EV) research field is the l (show more...)
EV-METRIC
50% (87th 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
Filtration SEC and UF Protein markers
EV: CD81/ Flotillin1/ CD63/ CD9
non-EV: Calreticulin Proteomics
no
Show all info
Study aim
Technical analysis comparing/optimizing EV-related methods
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
EV-producing cells
PC3
EV-harvesting Medium
EV-depleted medium
Preparation of EDS
Commercial EDS
Cell viability (%)
99
Cell count
7.50E+08
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
0.45µm > x > 0.22µm,
Other
Name other separation method
SEC and UF
Characterization: Protein analysis
Protein Concentration Method
microBCA
Western Blot
Antibody details provided?
Yes
Antibody dilution provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
Flotillin1/ CD63/ CD81
Not detected contaminants
Calreticulin
Flow cytometry
Type of Flow cytometry
NanoFCM
Hardware adaptation to ~100nm EV's
Please refer to the publication below: Zhu S, Ma L, Wang S, et al. Light-scattering detection below the level of single fluorescent molecules for high-resolution characterization of functional nanopar
Calibration bead size
0.2
Antibody details provided?
No
Detected EV-associated proteins
CD63/ CD9/ CD81
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Modus
Reported size (nm)
114.5
EV concentration
Yes
Particle yield
Yes, as number of particles per milliliter of starting sample 6.73E+07
Particle analysis: flow cytometry
Flow cytometer type
NanoFCM
Hardware adjustment
Please refer to the publication below: Zhu S, Ma L, Wang S, et al. Light-scattering detection below the level of single fluorescent molecules for high-resolution characterization of functional nanoparticles. ACS nano. 2014 Oct 28;8(10):10998-1006.
Calibration bead size
0.2
Report type
Modus
Reported size (nm)
59.25
EV concentration
Yes
|
||||||||
EV200093 | 5/10 | Homo sapiens | PC3 |
(d)(U)C Filtration DG |
Dong, Liang | 2021 | 50% | |
Study summaryFull title
All authors
Liang Dong, Richard C. Zieren, Kengo Horie, Chi‐Ju Kim, Emily Mallick, Yuezhou Jing, Mingxiao Feng, Morgan D. Kuczler, Jordan Green, Sarah R. Amend, Kenneth W. Witwer, Theo M. de Reijke, Yoon‐Kyoung Cho, Kenneth J. Pienta, Wei Xue
Journal
J Extracell Vesicles
Abstract
One of the challenges that restricts the evolving extracellular vesicle (EV) research field is the l (show more...)
EV-METRIC
50% (87th 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
Filtration DG Protein markers
EV: CD81/ Flotillin1
non-EV: None Proteomics
no
EV density (g/ml)
1.10-1.15
Show all info
Study aim
Technical analysis comparing/optimizing EV-related methods
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
EV-producing cells
PC3
EV-harvesting Medium
EV-depleted medium
Preparation of EDS
Commercial EDS
Cell viability (%)
99
Cell count
7.50E+08
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
Density gradient
Only used for validation of main results
Yes
Type
Continuous
Lowest density fraction
5%
Highest density fraction
60%
Total gradient volume, incl. sample (mL)
38
Sample volume (mL)
2
Orientation
Bottom-up
Rotor type
SW 32 Ti
Speed (g)
100000
Duration (min)
230
Fraction volume (mL)
4.75
Fraction processing
Centrifugation
Pelleting: volume per fraction
28
Pelleting: duration (min)
60
Pelleting: rotor type
Type 70 Ti
Pelleting: speed (g)
120000
Filtration steps
0.45µm > x > 0.22µm,
Characterization: Protein analysis
Protein Concentration Method
Not determined
Western Blot
Antibody details provided?
Yes
Antibody dilution provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
Flotillin1/ CD81
Flow cytometry
Type of Flow cytometry
NanoFCM
Hardware adaptation to ~100nm EV's
Please refer to the publication below: Zhu S, Ma L, Wang S, et al. Light-scattering detection below the level of single fluorescent molecules for high-resolution characterization of functional nanopar
Calibration bead size
0.2
Antibody details provided?
No
Characterization: Lipid analysis
No
Characterization: Particle analysis
Particle analysis: flow cytometry
Flow cytometer type
NanoFCM
Hardware adjustment
Please refer to the publication below: Zhu S, Ma L, Wang S, et al. Light-scattering detection below the level of single fluorescent molecules for high-resolution characterization of functional nanoparticles. ACS nano. 2014 Oct 28;8(10):10998-1006.
Calibration bead size
0.2
|
||||||||
EV200093 | 7/10 | Homo sapiens | Blood plasma |
(d)(U)C Filtration Total Exosome Isolation |
Dong, Liang | 2021 | 50% | |
Study summaryFull title
All authors
Liang Dong, Richard C. Zieren, Kengo Horie, Chi‐Ju Kim, Emily Mallick, Yuezhou Jing, Mingxiao Feng, Morgan D. Kuczler, Jordan Green, Sarah R. Amend, Kenneth W. Witwer, Theo M. de Reijke, Yoon‐Kyoung Cho, Kenneth J. Pienta, Wei Xue
Journal
J Extracell Vesicles
Abstract
One of the challenges that restricts the evolving extracellular vesicle (EV) research field is the l (show more...)
EV-METRIC
50% (83rd 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
(d)(U)C
Filtration Commercial method Protein markers
EV: CD81/ Flotillin1/ None/ CD63/ CD9
non-EV: ApoA1/ Calreticulin Proteomics
no
Show all info
Study aim
Technical analysis comparing/optimizing EV-related methods
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
0.45µm > x > 0.22µm,
Commercial kit
Total Exosome Isolation
Characterization: Protein analysis
Protein Concentration Method
BCA
Western Blot
Antibody details provided?
Yes
Antibody dilution provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
None
Not detected EV-associated proteins
CD81/ Flotillin1
Detected contaminants
ApoA1
Not detected contaminants
Calreticulin
Flow cytometry
Type of Flow cytometry
NanoFCM
Hardware adaptation to ~100nm EV's
Please refer to the publication below: Zhu S, Ma L, Wang S, et al. Light-scattering detection below the level of single fluorescent molecules for high-resolution characterization of functional nanopar
Calibration bead size
0.2
Antibody details provided?
No
Detected EV-associated proteins
None
Not detected EV-associated proteins
CD9/ CD63/ CD81
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Modus
Reported size (nm)
127.5
EV concentration
Yes
Particle yield
Yes, as number of particles per milliliter of starting sample 2.70E+11
Particle analysis: flow cytometry
Flow cytometer type
NanoFCM
Hardware adjustment
Please refer to the publication below: Zhu S, Ma L, Wang S, et al. Light-scattering detection below the level of single fluorescent molecules for high-resolution characterization of functional nanoparticles. ACS nano. 2014 Oct 28;8(10):10998-1006.
Calibration bead size
0.2
Report type
Modus
Reported size (nm)
61.75
EV concentration
Yes
|
||||||||
EV200093 | 8/10 | Homo sapiens | Blood plasma |
(d)(U)C Filtration Exodisc |
Dong, Liang | 2021 | 50% | |
Study summaryFull title
All authors
Liang Dong, Richard C. Zieren, Kengo Horie, Chi‐Ju Kim, Emily Mallick, Yuezhou Jing, Mingxiao Feng, Morgan D. Kuczler, Jordan Green, Sarah R. Amend, Kenneth W. Witwer, Theo M. de Reijke, Yoon‐Kyoung Cho, Kenneth J. Pienta, Wei Xue
Journal
J Extracell Vesicles
Abstract
One of the challenges that restricts the evolving extracellular vesicle (EV) research field is the l (show more...)
EV-METRIC
50% (83rd 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
(d)(U)C
Filtration Exodisc Protein markers
EV: CD81/ Flotillin1/ None/ CD63/ CD9
non-EV: ApoA1/ Calreticulin Proteomics
no
Show all info
Study aim
Technical analysis comparing/optimizing EV-related methods
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
0.45µm > x > 0.22µm,
Other
Name other separation method
Exodisc
Characterization: Protein analysis
Protein Concentration Method
BCA
Western Blot
Antibody details provided?
Yes
Antibody dilution provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
None
Not detected EV-associated proteins
CD81/ Flotillin1
Detected contaminants
ApoA1
Not detected contaminants
Calreticulin
Flow cytometry
Type of Flow cytometry
NanoFCM
Hardware adaptation to ~100nm EV's
Please refer to the publication below: Zhu S, Ma L, Wang S, et al. Light-scattering detection below the level of single fluorescent molecules for high-resolution characterization of functional nanopar
Calibration bead size
0.2
Antibody details provided?
No
Detected EV-associated proteins
None
Not detected EV-associated proteins
CD9/ CD63/ CD81
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Modus
Reported size (nm)
129.5
EV concentration
Yes
Particle yield
Yes, as number of particles per milliliter of starting sample 2.29E+11
Particle analysis: flow cytometry
Flow cytometer type
NanoFCM
Hardware adjustment
Please refer to the publication below: Zhu S, Ma L, Wang S, et al. Light-scattering detection below the level of single fluorescent molecules for high-resolution characterization of functional nanoparticles. ACS nano. 2014 Oct 28;8(10):10998-1006.
Calibration bead size
0.2
Report type
Modus
Reported size (nm)
62.25
EV concentration
Yes
|
||||||||
EV200093 | 9/10 | Homo sapiens | Blood plasma |
(d)(U)C Filtration SEC and UF |
Dong, Liang | 2021 | 50% | |
Study summaryFull title
All authors
Liang Dong, Richard C. Zieren, Kengo Horie, Chi‐Ju Kim, Emily Mallick, Yuezhou Jing, Mingxiao Feng, Morgan D. Kuczler, Jordan Green, Sarah R. Amend, Kenneth W. Witwer, Theo M. de Reijke, Yoon‐Kyoung Cho, Kenneth J. Pienta, Wei Xue
Journal
J Extracell Vesicles
Abstract
One of the challenges that restricts the evolving extracellular vesicle (EV) research field is the l (show more...)
EV-METRIC
50% (83rd 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
(d)(U)C
Filtration SEC and UF Protein markers
EV: CD81/ Flotillin1/ CD63/ CD9
non-EV: ApoA1/ Calreticulin Proteomics
no
Show all info
Study aim
Technical analysis comparing/optimizing EV-related methods
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
0.45µm > x > 0.22µm,
Other
Name other separation method
SEC and UF
Characterization: Protein analysis
Protein Concentration Method
BCA
Western Blot
Antibody details provided?
Yes
Antibody dilution provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
Flotillin1/ CD81
Detected contaminants
ApoA1
Not detected contaminants
Calreticulin
Flow cytometry
Type of Flow cytometry
NanoFCM
Hardware adaptation to ~100nm EV's
Please refer to the publication below: Zhu S, Ma L, Wang S, et al. Light-scattering detection below the level of single fluorescent molecules for high-resolution characterization of functional nanopar
Calibration bead size
0.2
Antibody details provided?
No
Detected EV-associated proteins
CD63/ CD9/ CD81
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Modus
Reported size (nm)
128.5
EV concentration
Yes
Particle yield
Yes, as number of particles per milliliter of starting sample 1.05E+11
Particle analysis: flow cytometry
Flow cytometer type
NanoFCM
Hardware adjustment
Please refer to the publication below: Zhu S, Ma L, Wang S, et al. Light-scattering detection below the level of single fluorescent molecules for high-resolution characterization of functional nanoparticles. ACS nano. 2014 Oct 28;8(10):10998-1006.
Calibration bead size
0.2
Report type
Modus
Reported size (nm)
58.25
EV concentration
Yes
|
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1 - 10 of 10 |
EV-TRACK ID | EV200093 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
species | Homo sapiens | |||||||||
sample type | Blood plasma | Cell culture | Blood plasma | Cell culture | Cell culture | Cell culture | Cell culture | Blood plasma | Blood plasma | Blood plasma |
cell type | NA | PC3 | NA | PC3 | PC3 | PC3 | PC3 | NA | NA | NA |
medium | NA | EV-depleted medium | NA | EV-depleted medium | EV-depleted medium | EV-depleted medium | EV-depleted medium | NA | NA | NA |
condition | Control condition | Control condition | Control condition | Control condition | Control condition | Control condition | Control condition | Control condition | Control condition | Control condition |
separation protocol | (d)(U)C Filtration DG | (d)(U)C Filtration | (d)(U)C Filtration | (d)(U)C Filtration Total Exosome Isolation | (d)(U)C Filtration Exodisc | (d)(U)C Filtration SEC and UF | (d)(U)C Filtration DG | (d)(U)C Filtration Total Exosome Isolation | (d)(U)C Filtration Exodisc | (d)(U)C Filtration SEC and UF |
Exp. nr. | 10 | 1 | 6 | 2 | 3 | 4 | 5 | 7 | 8 | 9 |
EV-METRIC % | 63 | 56 | 56 | 50 | 50 | 50 | 50 | 50 | 50 | 50 |