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Details | EV-TRACK ID | Experiment nr. | Species | Sample type | separation protocol | First author | Year | EV-METRIC |
---|---|---|---|---|---|---|---|---|
EV170028 | 1/1 | Homo sapiens | Synovial fluid |
DG (d)(U)C SEC |
Foers AD | 2017 | 100% | |
Study summaryFull title
All authors
Foers AD, Chatfield S, Dagley LF, Scicluna BJ, Webb AI, Cheng L, Hill AF, Wicks IP, Pang KC.
Journal
J Extracell Vesicles
Abstract
As a complex biological fluid, human synovial fluid (SF) presents challenges for extracellular vesic (show more...)
EV-METRIC
100% (85th 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
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
Synovial fluid
Sample origin
Arthritis
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
DG
(d)(U)C SEC Adj. k-factor
156.9 (pelleting) / 156.9 (washing)
Protein markers
EV: TSG101/ Rab27b/ Annexin1/ Syntenin/ HSP70/ Flotillin-1
non-EV: Fibronectin/ Albumin/ ApoA1 Proteomics
yes
Show all info
Study aim
New methodological development, Identification of content (omics approaches)
Sample
Species
Homo sapiens
Sample Type
Synovial fluid
Sample Condition
Arthritis
Separation Method
Differential ultracentrifugation
centrifugation steps
Between 100,000 g and 150,000 g
Obtain an EV pellet :
Yes
Pelleting: time(min)
90
Pelleting: rotor type
Type 70 Ti
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
156.9
Wash: time (min)
90
Wash: Rotor Type
Type 70 Ti
Wash: speed (g)
100000
Wash: adjusted k-factor
156.9
Density gradient
Density medium
Sucrose
Type
Discontinuous
Number of initial discontinuous layers
6
Lowest density fraction
0.25
Highest density fraction
2
Sample volume (mL)
0.65
Orientation
Top-down (sample migrates downwards)
Rotor type
SW 40 Ti
Speed (g)
202000
Duration (min)
1100
Fraction volume (mL)
1.05
Fraction processing
Centrifugation
Pelleting: volume per fraction
10
Pelleting: duration (min)
90
Pelleting: rotor type
Type 70.1Ti
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
122.2
Size-exclusion chromatography
Total column volume (mL)
320
Sample volume/column (mL)
13
Resin type
Sephacryl S-500 HR
Characterization: Protein analysis
Protein Concentration Method
BCA
Western Blot
Antibody details provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
Flotillin-1, HSP70, TSG101, Syntenin, Rab27b, Annexin1
Not detected contaminants
Albumin, ApoA1
Proteomics
Proteomics database
Yes
Characterization: Particle analysis
NTA
Report type
Mean
Reported size (nm)
x
EV concentration
Yes
Particle yield
x
EM
EM-type
Transmission-EM
Image type
Close-up, Wide-field
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