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You searched for: EV180017 (EV-TRACK ID)
Showing 1 - 5 of 5
Showing 1 - 5 of 5
Details | EV-TRACK ID | Experiment nr. | Species | Sample type | separation protocol | First author | Year | EV-METRIC |
---|---|---|---|---|---|---|---|---|
EV180017 | 1/5 | Homo sapiens | Cell culture supernatant | DG dUC Filtration Ultrafiltration UF |
Beghein E | 2018 | 63% | |
Study summaryFull title
All authors
Beghein E, Devriese D, Van Hoey E, Gettemans J
Journal
J Cell Sci
Abstract
Cancer cell-derived extracellular vesicles (EVs) are increasingly being recognized as genuine invasi (show more...)
EV-METRIC
63% (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
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 + dUC + Filtration + Ultrafiltration + UF
Protein markers
EV: TSG101/ CD63/ CD9/ MT1-MMP/ Flotillin-2
non-EV: Calnexin/ Golgin245/ PMP70/ CytochromeC Proteomics
yes
Show all info
Study aim
Biogenesis/cargo sorting, Identification of content (omics approaches)
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
Sample Condition
Control condition
EV-producing cells
MDAMB231
EV-harvesting Medium
Serum free medium
Separation Method
Differential ultracentrifugation
dUC: centrifugation steps
Below or equal to 800 g
Density gradient
Density medium
Iodixanol
Type
Discontinuous
Number of initial discontinuous layers
4
Lowest density fraction
0.05
Highest density fraction
0.4
Sample volume (mL)
0.5
Orientation
Top-down (sample migrates downwards)
Rotor type
SW 41 Ti
Speed (g)
100000
Duration (min)
1080
Fraction volume (mL)
1
Fraction processing
Centrifugation
Pelleting: volume per fraction
11
Pelleting: duration (min)
180
Pelleting: rotor type
SW 41 Ti
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
255.8
Filtration steps
0.45µm > x > 0.22µm, 0.22µm or 0.2µm
Ultra filtration
Cut-off size (kDa)
10
Membrane type
Regenerated cellulose
Characterization: Protein analysis
Protein Concentration Method
Not determined
Western Blot
Antibody details provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
CD9, CD63, Flotillin-2, TSG101, MT1-MMP
Not detected contaminants
PMP70, CytochromeC, Calnexin, Golgin245
Proteomics database
No
Characterization: Particle analysis
NTA
Report type
Mean
Reported size (nm)
131
EV concentration
Yes
Particle yield
4.95E+09 particles/million cells
|
||||||||
EV180017 | 2/5 | Homo sapiens | Cell culture supernatant | DG dUC Filtration Ultrafiltration UF |
Beghein E | 2018 | 62% | |
Study summaryFull title
All authors
Beghein E, Devriese D, Van Hoey E, Gettemans J
Journal
J Cell Sci
Abstract
Cancer cell-derived extracellular vesicles (EVs) are increasingly being recognized as genuine invasi (show more...)
EV-METRIC
62% (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
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
Expressing fascin nanobody
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
DG + dUC + Filtration + Ultrafiltration + UF
Protein markers
EV: CD63/ CD9
non-EV: None Proteomics
yes
Show all info
Study aim
Biogenesis/cargo sorting, Identification of content (omics approaches)
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
Sample Condition
Expressing fascin nanobody
EV-producing cells
MDAMB231
EV-harvesting Medium
Serum free medium
Separation Method
Differential ultracentrifugation
dUC: centrifugation steps
Below or equal to 800 g
Density gradient
Density medium
Iodixanol
Type
Discontinuous
Number of initial discontinuous layers
4
Lowest density fraction
0.05
Highest density fraction
0.4
Sample volume (mL)
0.5
Orientation
Top-down (sample migrates downwards)
Rotor type
SW 41 Ti
Speed (g)
100000
Duration (min)
1080
Fraction volume (mL)
1
Fraction processing
Centrifugation
Pelleting: volume per fraction
11
Pelleting: duration (min)
180
Pelleting: rotor type
SW 41 Ti
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
255.8
Filtration steps
0.45µm > x > 0.22µm, 0.22µm or 0.2µm
Ultra filtration
Cut-off size (kDa)
10
Membrane type
Regenerated cellulose
Characterization: Protein analysis
Protein Concentration Method
Not determined
Western Blot
Antibody details provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
CD9, CD63
Proteomics database
No
Characterization: Particle analysis
NTA
Report type
Mean
Reported size (nm)
142
EV concentration
Yes
Particle yield
2.25E+10 particles/million cells
|
||||||||
EV180017 | 5/5 | Homo sapiens | Cell culture supernatant | DG dUC Filtration Ultrafiltration UF |
Beghein E | 2018 | 62% | |
Study summaryFull title
All authors
Beghein E, Devriese D, Van Hoey E, Gettemans J
Journal
J Cell Sci
Abstract
Cancer cell-derived extracellular vesicles (EVs) are increasingly being recognized as genuine invasi (show more...)
EV-METRIC
62% (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
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
Expressing cortactin nanobody (NTA domain)
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
DG + dUC + Filtration + Ultrafiltration + UF
Protein markers
EV: CD63/ CD9
non-EV: None Proteomics
yes
Show all info
Study aim
Biogenesis/cargo sorting, Identification of content (omics approaches)
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
Sample Condition
Expressing cortactin nanobody (NTA domain)
EV-producing cells
MDAMB231
EV-harvesting Medium
Serum free medium
Separation Method
Differential ultracentrifugation
dUC: centrifugation steps
Below or equal to 800 g
Density gradient
Density medium
Iodixanol
Type
Discontinuous
Number of initial discontinuous layers
4
Lowest density fraction
0.05
Highest density fraction
0.4
Sample volume (mL)
0.5
Orientation
Top-down (sample migrates downwards)
Rotor type
SW 41 Ti
Speed (g)
100000
Duration (min)
1080
Fraction volume (mL)
1
Fraction processing
Centrifugation
Pelleting: volume per fraction
11
Pelleting: duration (min)
180
Pelleting: rotor type
SW 41 Ti
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
255.8
Filtration steps
0.45µm > x > 0.22µm, 0.22µm or 0.2µm
Ultra filtration
Cut-off size (kDa)
10
Membrane type
Regenerated cellulose
Characterization: Protein analysis
Protein Concentration Method
Not determined
Western Blot
Antibody details provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
CD9, CD63
Proteomics database
No
Characterization: Particle analysis
NTA
Report type
Mean
Reported size (nm)
133
EV concentration
Yes
Particle yield
1.16E+09 particles/million cells
|
||||||||
EV180017 | 3/5 | Homo sapiens | Cell culture supernatant | DG dUC Filtration Ultrafiltration UF |
Beghein E | 2018 | 50% | |
Study summaryFull title
All authors
Beghein E, Devriese D, Van Hoey E, Gettemans J
Journal
J Cell Sci
Abstract
Cancer cell-derived extracellular vesicles (EVs) are increasingly being recognized as genuine invasi (show more...)
EV-METRIC
50% (85th 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
Sample origin
Expressing cortactin nanobody (SH3 domain)
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
DG + dUC + Filtration + Ultrafiltration + UF
Protein markers
EV: CD63/ CD9
non-EV: None Proteomics
no
Show all info
Study aim
Biogenesis/cargo sorting, Identification of content (omics approaches)
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
Sample Condition
Expressing cortactin nanobody (SH3 domain)
EV-producing cells
MDAMB231
EV-harvesting Medium
Serum free medium
Separation Method
Differential ultracentrifugation
dUC: centrifugation steps
Below or equal to 800 g
Density gradient
Density medium
Iodixanol
Type
Discontinuous
Number of initial discontinuous layers
4
Lowest density fraction
0.05
Highest density fraction
0.4
Sample volume (mL)
0.5
Orientation
Top-down (sample migrates downwards)
Rotor type
SW 41 Ti
Speed (g)
100000
Duration (min)
1080
Fraction volume (mL)
1
Fraction processing
Centrifugation
Pelleting: volume per fraction
11
Pelleting: duration (min)
180
Pelleting: rotor type
SW 41 Ti
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
255.8
Filtration steps
0.45µm > x > 0.22µm, 0.22µm or 0.2µm
Ultra filtration
Cut-off size (kDa)
10
Membrane type
Regenerated cellulose
Characterization: Protein analysis
Protein Concentration Method
Not determined
Western Blot
Antibody details provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
CD9, CD63
Characterization: Particle analysis
NTA
Report type
Mean
Reported size (nm)
133
EV concentration
Yes
Particle yield
9.79E+09 particles/million cells
|
||||||||
EV180017 | 4/5 | Homo sapiens | Cell culture supernatant | DG dUC Filtration Ultrafiltration UF |
Beghein E | 2018 | 50% | |
Study summaryFull title
All authors
Beghein E, Devriese D, Van Hoey E, Gettemans J
Journal
J Cell Sci
Abstract
Cancer cell-derived extracellular vesicles (EVs) are increasingly being recognized as genuine invasi (show more...)
EV-METRIC
50% (85th 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
Sample origin
Expressing gelsolin nanobody
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
DG + dUC + Filtration + Ultrafiltration + UF
Protein markers
EV: CD63/ CD9
non-EV: None Proteomics
no
Show all info
Study aim
Biogenesis/cargo sorting, Identification of content (omics approaches)
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
Sample Condition
Expressing gelsolin nanobody
EV-producing cells
MDAMB231
EV-harvesting Medium
Serum free medium
Separation Method
Differential ultracentrifugation
dUC: centrifugation steps
Below or equal to 800 g
Density gradient
Density medium
Iodixanol
Type
Discontinuous
Number of initial discontinuous layers
4
Lowest density fraction
0.05
Highest density fraction
0.4
Sample volume (mL)
0.5
Orientation
Top-down (sample migrates downwards)
Rotor type
SW 41 Ti
Speed (g)
100000
Duration (min)
1080
Fraction volume (mL)
1
Fraction processing
Centrifugation
Pelleting: volume per fraction
11
Pelleting: duration (min)
180
Pelleting: rotor type
SW 41 Ti
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
255.8
Filtration steps
0.45µm > x > 0.22µm, 0.22µm or 0.2µm
Ultra filtration
Cut-off size (kDa)
10
Membrane type
Regenerated cellulose
Characterization: Protein analysis
Protein Concentration Method
Not determined
Western Blot
Antibody details provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
CD9, CD63
Characterization: Particle analysis
NTA
Report type
Mean
Reported size (nm)
128
EV concentration
Yes
Particle yield
1.97E+09 particles/million cells
|
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1 - 5 of 5 |
EV-TRACK ID | EV180017 | ||||
---|---|---|---|---|---|
species | Homo sapiens | ||||
sample type | Cell culture | ||||
cell type | MDAMB231 | ||||
condition | Control condition | Expressing fascin nanobody | Expressing cortactin nanobody (NTA domain) | Expressing cortactin nanobody (SH3 domain) | Expressing gelsolin nanobody |
separation protocol | DG dUC Filtration Ultrafiltration UF | DG dUC Filtration Ultrafiltration UF | DG dUC Filtration Ultrafiltration UF | DG dUC Filtration Ultrafiltration UF | DG dUC Filtration Ultrafiltration UF |
Exp. nr. | 1 | 2 | 5 | 3 | 4 |
EV-METRIC % | 63 | 62 | 62 | 50 | 50 |