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You searched for: EV170016 (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 |
---|---|---|---|---|---|---|---|---|
EV170016 | 1/2 | Mus musculus | adipose tissue-derived macrophages |
DG (d)(U)C Filtration |
Ying, Wei | 2017 | 88% | |
Study summaryFull title
All authors
Ying W, Riopel M, Bandyopadhyay G, Dong Y, Birmingham A, Seo JB, Ofrecio JM, Wollam J, Hernandez-Carretero A, Fu W, Li P, Olefsky JM
Journal
Cell
Abstract
MiRNAs are regulatory molecules that can be packaged into exosomes and secreted from cells. Here, we (show more...)
EV-METRIC
88% (98th 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
Obesitas
Focus vesicles
exosome
Separation protocol
Separation protocol
DG
(d)(U)C Filtration Adj. k-factor
256 (pelleting) / 256 (washing)
Protein markers
EV: TSG101/ HSP70/ Syntenin1/ CD63/ CD9
non-EV: Grp94 Proteomics
no
Show all info
Study aim
Function, Mechanism of uptake/transfer
Sample
Species
Mus musculus
Sample Type
Cell culture supernatant
EV-producing cells
adipose tissue-derived macrophages
EV-harvesting Medium
EV-depleted serum
Preparation of EDS
Commercial EDS
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 800 g and 10,000 g
Between 100,000 g and 150,000 g Pelleting performed
Yes
Pelleting: time(min)
240-360
Pelleting: rotor type
SW 32 Ti
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
256.0
Wash: time (min)
20
Wash: Rotor Type
SW 32 Ti
Wash: speed (g)
100000
Wash: adjusted k-factor
256.0
Density gradient
Only used for validation of main results
Yes
Density medium
156.9
Type
Discontinuous
Lowest density fraction
0.1
Highest density fraction
0.3
Sample volume (mL)
0.3
Orientation
Bottom-up (sample migrates upwards)
Rotor type
Type 70 Ti
Speed (g)
350000
Duration (min)
60
Fraction volume (mL)
2.4
Fraction processing
Centrifugation
Pelleting: volume per fraction
2
Pelleting: duration (min)
90
Pelleting: rotor type
Type 70 Ti
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
156.9
Pelleting-wash: volume per pellet (mL)
24
Pelleting-wash: duration (min)
30
Pelleting-wash: rotor type
156.9
Pelleting-wash: speed (g)
Type 70 Ti
Pelleting-wash: adjusted k-factor
156.9
Filtration steps
0.22µm or 0.2µm
EV-subtype
Distinction between multiple subtypes
1.13-1.15 g/ml
Used subtypes
Yes
Characterization: Protein analysis
Protein Concentration Method
DC protein assay
Protein Yield (µg)
9-May
Western Blot
Antibody details provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
CD9, CD63, HSP70, TSG101, Syntenin1
Not detected contaminants
Grp94
Characterization: RNA analysis
Database
No
Proteinase treatment
No
RNAse treatment
No
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Size range/distribution
Reported size (nm)
30-150
EM
EM-type
Transmission-EM
Image type
Wide-field
Report size (nm)
50-200 nm
Extra information
Full UC and density gradient protocols not in original article
|
||||||||
EV170016 | 2/2 | Mus musculus | adipose tissue-derived macrophages |
(d)(U)C Filtration |
Ying, Wei | 2017 | 55% | |
Study summaryFull title
All authors
Ying W, Riopel M, Bandyopadhyay G, Dong Y, Birmingham A, Seo JB, Ofrecio JM, Wollam J, Hernandez-Carretero A, Fu W, Li P, Olefsky JM
Journal
Cell
Abstract
MiRNAs are regulatory molecules that can be packaged into exosomes and secreted from cells. Here, we (show more...)
EV-METRIC
55% (89th 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
(d)(U)C
Filtration Adj. k-factor
256 (pelleting) / 256 (washing)
Protein markers
EV: TSG101/ HSP70/ Syntenin1/ CD63/ CD9
non-EV: Grp94 Proteomics
no
Show all info
Study aim
Function, Mechanism of uptake/transfer
Sample
Species
Mus musculus
Sample Type
Cell culture supernatant
EV-producing cells
adipose tissue-derived macrophages
EV-harvesting Medium
EV-depleted serum
Preparation of EDS
Commercial EDS
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Between 800 g and 10,000 g
Between 100,000 g and 150,000 g Pelleting performed
Yes
Pelleting: time(min)
240-360
Pelleting: rotor type
SW 32 Ti
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
256.0
Wash: time (min)
20
Wash: Rotor Type
SW 32 Ti
Wash: speed (g)
100000
Wash: adjusted k-factor
256.0
Filtration steps
0.22µm or 0.2µm
EV-subtype
Distinction between multiple subtypes
1.13-1.15 g/ml
Used subtypes
Yes
Characterization: Protein analysis
Protein Concentration Method
DC protein assay
Protein Yield (µg)
9-May
Western Blot
Antibody details provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
CD9, CD63, HSP70, TSG101, Syntenin1
Not detected contaminants
Grp94
Characterization: RNA analysis
Database
No
Proteinase treatment
No
RNAse treatment
No
Characterization: Lipid analysis
No
Extra information
Full UC protocol not in original article
|
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1 - 2 of 2 |
EV-TRACK ID | EV170016 | |
---|---|---|
species | Mus musculus | |
sample type | Cell culture | |
cell type | adipose tissue-derived macrophages | |
condition | Obesitas | Control condition |
separation protocol | DG (d)(U)C Filtration | (d)(U)C Filtration |
Exp. nr. | 1 | 2 |
EV-METRIC % | 88 | 55 |