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You searched for: EV170030 (EV-TRACK ID)
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Details | EV-TRACK ID | Experiment nr. | Species | Sample type | Separation protocol | First author | Year | EV-METRIC |
---|---|---|---|---|---|---|---|---|
EV170030 | 1/1 | Mus musculus | primary bone marrow dendritic cells |
DG (d)(U)C |
Driedonks, Tom A. P. | 2018 | 100% | |
Study summaryFull title
All authors
Driedonks TAP, van der Grein SG, Ariyurek Y, Buermans HPJ, Jekel H, Chow FWN, Wauben MHM, Buck AH, 't Hoen PAC, Nolte-'t Hoen ENM
Journal
Cell Mol Life Sci
Abstract
The release and uptake of nano-sized extracellular vesicles (EV) is a highly conserved means of inte (show more...)
EV-METRIC
100% (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
Control condition
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
DG
(d)(U)C Adj. k-factor
253.9 (pelleting)
Protein markers
EV: MHC2/ CD63/ CD9/ Galectin-3
non-EV: beta-actin Proteomics
no
Show all info
Study aim
Biomarker, Biogenesis/cargo sorting, Identification of content (omics approaches)
Sample
Species
Mus musculus
Sample Type
Cell culture supernatant
EV-producing cells
primary bone marrow dendritic cells
EV-harvesting Medium
EV-depleted serum
Preparation of EDS
overnight (16h) at >=100,000g
Cell viability (%)
NA
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: time(min)
65
Pelleting: rotor type
SW 28
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
253.9
Density gradient
Type
Continuous
Lowest density fraction
0.4M
Highest density fraction
2.5M
Sample volume (mL)
0.15
Orientation
Bottom-up (sample migrates upwards)
Rotor type
SW 40 Ti
Speed (g)
192000
Duration (min)
900-1080
Fraction volume (mL)
1
Fraction processing
Centrifugation
Pelleting: volume per fraction
1
Pelleting: duration (min)
65
Pelleting: rotor type
SW 40 Ti
Pelleting: speed (g)
192000
Pelleting: adjusted k-factor
144.0
Characterization: Protein analysis
Protein Concentration Method
Not determined
Western Blot
Detected EV-associated proteins
CD9, CD63, MHC2, Galectin-3
Not detected contaminants
beta-actin
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)
100 - 200
EV concentration
Yes
Particle analysis: flow cytometry
Flow cytometer type
BD Influx
Hardware adjustment
see van der Vlist et al. 2012 Nature Protocols;and Nolte-'t Hoen 2012 Nanomedicine
Calibration bead size
0.1,0.2
EV concentration
Yes
EM
EM-type
Transmission-EM
Image type
Close-up, Wide-field
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EV-TRACK ID | EV170030 |
---|---|
species | Mus musculus |
sample type | Cell culture |
cell type | primary bone marrow dendritic cells |
condition | Control condition |
separation protocol | DG (d)(U)C |
Exp. nr. | 1 |
EV-METRIC % | 100 |