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You searched for: EV180042 (EV-TRACK ID)
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Details | EV-TRACK ID | Experiment nr. | Species | Sample type | Separation protocol | First author | Year | EV-METRIC |
---|---|---|---|---|---|---|---|---|
EV180042 | 1/1 | Mus musculus | RAW264.7,mouse primary osteoclasts | (d)(U)C | Ikebuchi, Yuki | 2018 | 55% | |
Study summaryFull title
All authors
Ikebuchi Y, Aoki S, Honma M, Hayashi M, Sugamori Y, Khan M, Kariya Y, Kato G, Tabata Y, Penninger JM, Udagawa N, Aoki K, Suzuki H.
Journal
Nature
Abstract
Receptor activator of nuclear factor-kappa B (RANK) ligand (RANKL) binds RANK on the surface of oste (show more...)
EV-METRIC
55% (88th 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
RANKL-stimulated
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
(d)(U)C
Adj. k-factor
209.7 (pelleting) / 209.7 (washing)
Protein markers
EV: CD81/ CD63/ CD9/ RANK
non-EV: Calnexin Proteomics
yes
Show all info
Study aim
Function, Identification of content (omics approaches)
Sample
Species
Mus musculus
Sample Type
Cell culture supernatant
EV-producing cells
RAW264.7,mouse primary osteoclasts
EV-harvesting Medium
EV-depleted serum
Preparation of EDS
>=18h 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)
60
Pelleting: rotor type
Type 45 Ti
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
209.7
Wash: time (min)
60
Wash: Rotor Type
Type 45 Ti
Wash: speed (g)
100000
Wash: adjusted k-factor
209.7
Characterization: Protein analysis
Protein Concentration Method
BCA
Protein Yield (µg)
2.5-3.5
Western Blot
Antibody details provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
CD9, CD63, CD81
Not detected contaminants
Calnexin
ELISA
Antibody details provided?
Yes
Lysis buffer provided?
Yes
Proteomics database
Yes
Characterization: Lipid analysis
No
Characterization: Particle analysis
EM
EM-type
Transmission-EM
Image type
Close-up
Extra information
Antibody details for Western blot: rabbit anti-CD9 (Abcam, CatNo. ab92726, clone EPR2949, LotNo. GR260186-10, 1:1000), rat anti-CD63 (MBL, CatNo. D263-3, clone R5G2, LotNo. 014, 1:500), hamster anti-CD81 (Bio-Rad, CatNo. MCA1846GA, clone Eat2, LotNo. 0515, 1:500), rabbit anti-Calnexin (Abcam, CatNo. ab22595, polyclonal, LotNo. GR86850-1, 1:500)
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EV-TRACK ID | EV180042 |
---|---|
species | Mus musculus |
sample type | Cell culture |
cell type | RAW264.7 mouse primary osteoclasts |
condition | RANKL-stimulated |
separation protocol | (d)(U)C |
Exp. nr. | 1 |
EV-METRIC % | 55 |