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You searched for: EV190103 (EV-TRACK ID)
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Details | EV-TRACK ID | Experiment nr. | Species | Sample type | separation protocol | First author | Year | EV-METRIC |
---|---|---|---|---|---|---|---|---|
EV190103 | 1/1 | Homo sapiens | Cell culture supernatant |
DG (d)(U)C Dynabeads CD9, CD63, CD81 |
Xiaoju Zhou | 2020 | 100% | |
Study summaryFull title
All authors
Xiaoju Zhou, Brooke A Brown, Amanda P Siegel, Mohamed El Masry, Xuyao Zeng, Woran Song, Amitava Das, Puneet Khandelwal, Andrew Clark, Kanhaiya Singh, Poornachander R Guda, Mahadeo Gorain, Lava Timsina, Yi Xuan, Stephen C Jacobson, Milos V Novotny, Sashwati Roy, Mangilal Agarwal, Robert J Lee, Chandan K Sen, David E Clemmer, Subhadip Ghatak
Journal
ACS Nano
Abstract
Bidirectional cell-cell communication involving exosome-borne cargo such as miRNA, has emerged as a (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
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
Cell Name
HaCaT
Sample origin
Control condition
Focus vesicles
exosome
Separation protocol
Separation protocol
DG
(d)(U)C Dynabeads CD9, CD63, CD81 Protein markers
EV: TSG101/ Alix
non-EV: Proteomics
no
EV density (g/ml)
1.16
Show all info
Study aim
Function/Biogenesis/cargo sorting/Mechanism of uptake/transfer
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
Sample Condition
Control condition
EV-producing cells
HaCaT
EV-harvesting Medium
EV-depleted medium
Preparation of EDS
Commercial EDS
Cell viability
Yes
Cell viability (%)
Yes
Separation Method
Differential ultracentrifugation
centrifugation steps
Between 800 g and 10,000 g
Between 10,000 g and 50,000 g Equal to or above 150,000 g Obtain an EV pellet :
Yes
Pelleting: time(min)
120
Pelleting: rotor type
TLA-120.2
Pelleting: speed (g)
245000
Wash: volume per pellet (ml)
0.5
Wash: time (min)
120
Wash: Rotor Type
TLA-120.2
Wash: speed (g)
245000
Density gradient
Only used for validation of main results
Yes
Density medium
Sucrose
Type
Discontinuous
Number of initial discontinuous layers
6
Lowest density fraction
0.4M
Highest density fraction
2.5M
Total gradient volume, incl. sample (mL)
1.05
Sample volume (mL)
0.05
Orientation
Top-down
Rotor type
TLA-120.2
Speed (g)
100000
Duration (min)
1080
Fraction volume (mL)
0.1
Fraction processing
Centrifugation
Pelleting: volume per fraction
1
Pelleting: duration (min)
120
Pelleting: rotor type
TLA-120.2
Pelleting: speed (g)
245000
Commercial kit
Dynabeads CD9, CD63, CD81
Characterization: Protein analysis
Protein Concentration Method
microBCA
Western Blot
Detected EV-associated proteins
TSG101/ Alix/ FLOT1/ HSP90
Detected contaminants
Prohibitin
Not detected contaminants
GM130
Flow cytometry specific beads
Detected EV-associated proteins
TSG101
Other 1
Fluorescent anisotropy
Detected EV-associated proteins
TSG101
Characterization: Particle analysis
NTA
Report type
Modus
Reported size (nm)
102
EV concentration
Yes
Particle yield
Yes, as number of particles per milliliter of starting sample 3.20E+07
EM
EM-type
Scanning-EM
Image type
Wide-field
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