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You searched for: EV200042 (EV-TRACK ID)
Showing 1 - 2 of 2
Showing 1 - 2 of 2
Details | EV-TRACK ID | Experiment nr. | Species | Sample type | Separation protocol | First author | Year | EV-METRIC |
---|---|---|---|---|---|---|---|---|
EV200042 | 1/2 | Mus musculus | Bone marrow-derived macrophages (BMDMs) |
DG (d)(U)C DC Filtration |
Laura Bouchareychas | 2020 | 75% | |
Study summaryFull title
All authors
Laura Bouchareychas, Phat Duong, Sergio Covarrubias, Eric Alsop, Tuan Anh Phu, Allen Chung, Michael Gomes, David Wong, Bessie Meechoovet, Allyson Capili, Ryo Yamamoto, Hiromitsu Nakauchi, Michael T McManus, Susan Carpenter, Kendall Van Keuren-Jensen, Robert L Raffai
Journal
Cell Rep
Abstract
Developing strategies that promote the resolution of vascular inflammation and atherosclerosis remai (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
Cell culture supernatant
Sample origin
Control condition
Focus vesicles
exosome
Separation protocol
Separation protocol
DG
(d)(U)C DC Filtration Protein markers
EV: Alix/ Flotillin1/ CD9
non-EV: Calnexin/ GM130 Proteomics
no
EV density (g/ml)
1.09
Show all info
Study aim
Function/Identification of content (omics approaches)
Sample
Species
Mus musculus
Sample Type
Cell culture supernatant
EV-producing cells
Bone marrow-derived macrophages (BMDMs)
EV-harvesting Medium
EV-depleted medium
Preparation of EDS
>=18h at >= 100,000g
Cell viability (%)
95
Cell count
5.00E+06
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
No
Density gradient
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
SW 40 Ti
Speed (g)
100000
Duration (min)
1080
Fraction volume (mL)
1
Fraction processing
None
Filtration steps
0.22µm or 0.2µm
Density cushion
Density medium
Iodixanol
Characterization: Protein analysis
Protein Concentration Method
Fluorometric assay (e.g. Qubit, NanoOrange,...)
Western Blot
Antibody details provided?
Yes
Antibody dilution provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
Flotillin1/ CD9/ Alix
Not detected contaminants
Calnexin/ GM130
Characterization: RNA analysis
RNA analysis
Type
RNA sequencing
Database
Yes
Proteinase treatment
No
RNAse treatment
Yes
Moment of RNAse treatment
After
RNAse type
Other;RNase A/T1 Mix
RNAse concentration
0.4
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Size range/distribution
Reported size (nm)
75.66
EV concentration
Yes
Particle yield
Yes, as number of particles per million cells 2.70E+09
|
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EV200042 | 2/2 | Mus musculus | Immortalized bone marrow-derived macrophages (iBMDMs) |
DG (d)(U)C DC Filtration |
Laura Bouchareychas | 2020 | 63% | |
Study summaryFull title
All authors
Laura Bouchareychas, Phat Duong, Sergio Covarrubias, Eric Alsop, Tuan Anh Phu, Allen Chung, Michael Gomes, David Wong, Bessie Meechoovet, Allyson Capili, Ryo Yamamoto, Hiromitsu Nakauchi, Michael T McManus, Susan Carpenter, Kendall Van Keuren-Jensen, Robert L Raffai
Journal
Cell Rep
Abstract
Developing strategies that promote the resolution of vascular inflammation and atherosclerosis remai (show more...)
EV-METRIC
63% (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
Cell culture supernatant
Sample origin
genetically modified cell line
Focus vesicles
exosome
Separation protocol
Separation protocol
DG
(d)(U)C DC Filtration Protein markers
EV: Alix/ Flotillin1
non-EV: Calnexin/ GM130 Proteomics
no
EV density (g/ml)
1.09
Show all info
Study aim
Function/Identification of content (omics approaches)
Sample
Species
Mus musculus
Sample Type
Cell culture supernatant
EV-producing cells
Immortalized bone marrow-derived macrophages (iBMDMs)
EV-harvesting Medium
EV-depleted medium
Preparation of EDS
>=18h at >= 100,000g
Cell viability (%)
95
Cell count
3.50E+07
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
No
Density gradient
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
SW 40 Ti
Speed (g)
100000
Duration (min)
1080
Fraction volume (mL)
1
Fraction processing
None
Filtration steps
0.22µm or 0.2µm
Density cushion
Density medium
Iodixanol
Characterization: Protein analysis
Protein Concentration Method
Fluorometric assay (e.g. Qubit, NanoOrange,...)
Western Blot
Antibody details provided?
Yes
Antibody dilution provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
Flotillin1/ Alix
Not detected contaminants
Calnexin/ GM130
Characterization: RNA analysis
RNA analysis
Type
(RT)(q)PCR
Database
No
Proteinase treatment
No
RNAse treatment
Yes
Moment of RNAse treatment
After
RNAse type
Other;RNase A/T1 Mix
RNAse concentration
0.4
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Size range/distribution
Reported size (nm)
74.58
EV concentration
Yes
Particle yield
Yes, as number of particles per million cells 2.40E+09
|
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1 - 2 of 2 |
EV-TRACK ID | EV200042 | |
---|---|---|
species | Mus musculus | |
sample type | Cell culture | |
cell type | Bone marrow-derived macrophages (BMDMs) | Immortalized bone marrow-derived macrophages (iBMDMs) |
condition | Control condition | genetically modified cell line |
separation protocol | DG (d)(U)C DC Filtration | DG (d)(U)C DC Filtration |
Exp. nr. | 1 | 2 |
EV-METRIC % | 75 | 63 |