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You searched for: EV210021 (EV-TRACK ID)
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Showing 1 - 2 of 2
Details | EV-TRACK ID | Experiment nr. | Species | Sample type | Separation protocol | First author | Year | EV-METRIC |
---|---|---|---|---|---|---|---|---|
EV210021 | 1/2 | Rattus norvegicus | Primary rat hepatocytes |
(d)(U)C Filtration DG |
Mleczko, J.E. | 2021 | 89% | |
Study summaryFull title
All authors
J.E. Mleczko, F. Royo, I. Samuelson, M. Clos-Garcia, C. Williams, D. Cabrera, M. Azparren-Angulo, E. Gonzalez, C. Garcia-Vallicrosa, S. Carobbio, S. Rodriguez-Cuenca, M. Azkargorta, S. van Liempd, F. Elortza, A. Vidal-Puig, S. Mora, J.M. Falcon-Perez
Journal
Journal of Extracellular Biology
Abstract
The composition of extracellular vesicles (EVs) is altered in many pathological condi-tions, and the (show more...)
EV-METRIC
89% (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. 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
Obese hepatocytes
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
(Differential) (ultra)centrifugation
Filtration Density gradient Protein markers
EV: TSG101/ CD63/ CD81/ HSP90/ Alix/ Flotillin1/ HSP70
non-EV: GRP78/ COXIV/ Parp Proteomics
yes
Show all info
Study aim
Function/Identification of content (omics approaches)
Sample
Species
Rattus norvegicus
Sample Type
Cell culture supernatant
EV-producing cells
Primary rat hepatocytes
EV-harvesting Medium
EV-depleted medium
Preparation of EDS
overnight (16h) at >=100,000g
Cell count
300000000
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 Between 50,000 g and 100,000 g Pelleting performed
Yes
Pelleting: rotor type
Type 45 Ti
Pelleting: speed (g)
100000
Wash: volume per pellet (ml)
45
Wash: time (min)
90
Wash: Rotor Type
Type 45 Ti
Wash: speed (g)
100000
Density gradient
Only used for validation of main results
Yes
Type
Discontinuous
Number of initial discontinuous layers
2
Lowest density fraction
5%
Highest density fraction
40%
Total gradient volume, incl. sample (mL)
16,5
Sample volume (mL)
5,5
Orientation
Top-down
Speed (g)
100000
Duration (min)
160
Fraction volume (mL)
1
Fraction processing
Centrifugation
Pelleting: volume per fraction
Not repo
Pelleting: duration (min)
60
Pelleting: rotor type
Not reported
Pelleting: speed (g)
100000
Filtration steps
0.22µm or 0.2µm
Characterization: Protein analysis
Protein Concentration Method
Bradford
Western Blot
Detected EV-associated proteins
Flotillin1/ CD63/ HSP90/ TSG101/ HSP70/ Alix/ CD81
Not detected EV-associated proteins
HSP90/ HSP70/ CD81/ Flotillin1/ TSG101/ CD63/ Alix
Detected contaminants
COXIV/ GRP78/ Parp
Not detected contaminants
COXIV/ GRP78/ Parp
Proteomics database
No
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Size range/distribution
Reported size (nm)
50-400
EV concentration
Yes
Particle yield
as number of particles per million cells: 2
EM
EM-type
Cryo-EM
Image type
Close-up
|
||||||||
EV210021 | 2/2 | Rattus norvegicus | Primary rat hepatocytes |
(d)(U)C Filtration |
Mleczko, J.E. | 2021 | 89% | |
Study summaryFull title
All authors
J.E. Mleczko, F. Royo, I. Samuelson, M. Clos-Garcia, C. Williams, D. Cabrera, M. Azparren-Angulo, E. Gonzalez, C. Garcia-Vallicrosa, S. Carobbio, S. Rodriguez-Cuenca, M. Azkargorta, S. van Liempd, F. Elortza, A. Vidal-Puig, S. Mora, J.M. Falcon-Perez
Journal
Journal of Extracellular Biology
Abstract
The composition of extracellular vesicles (EVs) is altered in many pathological condi-tions, and the (show more...)
EV-METRIC
89% (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. 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
Lean hepatocytes
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
(Differential) (ultra)centrifugation
Filtration Protein markers
EV: TSG101/ CD63/ CD81/ HSP90/ Alix/ Flotillin1/ HSP70
non-EV: Grp78/ GRP78/ COXIV/ Parp Proteomics
yes
Show all info
Study aim
Function/Identification of content (omics approaches)
Sample
Species
Rattus norvegicus
Sample Type
Cell culture supernatant
EV-producing cells
Primary rat hepatocytes
EV-harvesting Medium
EV-depleted medium
Preparation of EDS
overnight (16h) at >=100,000g
Cell count
300000000
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: rotor type
Type 45 Ti
Pelleting: speed (g)
100000
Wash: volume per pellet (ml)
45
Wash: time (min)
90
Wash: Rotor Type
Type 45 Ti
Wash: speed (g)
100000
Density gradient
Only used for validation of main results
Yes
Type
Discontinuous
Number of initial discontinuous layers
2
Lowest density fraction
5%
Highest density fraction
40%
Total gradient volume, incl. sample (mL)
16,5
Sample volume (mL)
5,5
Orientation
Top-down
Speed (g)
100000
Duration (min)
160
Fraction volume (mL)
1
Fraction processing
Centrifugation
Pelleting: volume per fraction
Not repo
Pelleting: duration (min)
60
Pelleting: rotor type
Not reported
Pelleting: speed (g)
100000
Filtration steps
0.22µm or 0.2µm
Characterization: Protein analysis
Protein Concentration Method
Bradford
Western Blot
Detected EV-associated proteins
Flotillin1/ CD63/ HSP90/ TSG101/ HSP70/ Alix/ CD81
Not detected EV-associated proteins
HSP90/ HSP70/ CD81/ Flotillin1/ TSG101/ CD63/ Alix
Detected contaminants
COXIV/ GRP78/ Parp
Not detected contaminants
COXIV/ Grp78/ Parp
Proteomics database
No
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Size range/distribution
Reported size (nm)
50-400
EV concentration
Yes
Particle yield
as number of particles per million cells: 1,8
EM
EM-type
Cryo-EM
Image type
Close-up
|
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1 - 2 of 2 |
EV-TRACK ID | EV210021 | |
---|---|---|
species | Rattus norvegicus | |
sample type | Cell culture | |
cell type | Primary rat hepatocytes | |
condition | Obese hepatocytes | Lean hepatocytes |
separation protocol | dUC/ Filtration/ Density gradient | dUC/ Filtration |
Exp. nr. | 1 | 2 |
EV-METRIC % | 89 | 89 |