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You searched for: EV210279 (EV-TRACK ID)

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Experiment number
  • If needed, multiple experiments were identified in a single publication based on differing sample types, separation protocols and/or vesicle types of interest.
Species
  • Species of origin of the EVs.
Separation protocol
  • Gives a short, non-chronological overview of the different steps of the separation protocol.
    • (d)(U)C = (differential) (ultra)centrifugation
    • DG = density gradient
    • UF = ultrafiltration
    • SEC = size-exclusion chromatography
    • IAF = immuno-affinity capture
Details EV-TRACK code Experiment nr. Species Sample type Separation protocol First author Year EV-METRIC
EV210279 1/2 Homo sapiens Cell culture supernatant (d)(U)C
DG
Libregts SFWM 2018 63%

Study summary

Full title
All authors
Libregts SFWM, Arkesteijn GJA, Németh A, Nolte-'t Hoen ENM, Wauben MHM
Journal
J Thromb Haemost
Abstract
Essentials Extracellular vesicles (EVs) in biological fluids are promising biomarkers for disease. F (show more...)Essentials Extracellular vesicles (EVs) in biological fluids are promising biomarkers for disease. Fluorescence-based flow cytometric analysis is suitable to detect low abundant EV subsets. Particles of non-interest can induce false-positive light scatter and fluorescent signals. Interference of particles of non-interest can be monitored by analyzing serial dilutions. (hide)
EV-METRIC
63% (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 data
Sample type
Cell culture supernatant
Sample origin
Control condition
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
  • Gives a short, non-chronological overview of the
    different steps of the separation protocol.
    • dUC = (Differential) (ultra)centrifugation
    • DG = density gradient
    • UF = ultrafiltration
    • SEC = size-exclusion chromatography
    • IAF = immuno-affinity capture
(Differential) (ultra)centrifugation
Density gradient
Protein markers
EV: EGFR/ CD9/ CD142
non-EV: None
Proteomics
no
Show all info
Study aim
Technical analysis comparing/optimizing EV-related methods
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
EV-producing cells
A431
EV-harvesting Medium
EV-depleted medium
Preparation of EDS
overnight (16h) at >=100,000g
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: rotor type
SW28 or SW32
Pelleting: speed (g)
100000
Density gradient
Type
Continuous
Lowest density fraction
0.4M
Highest density fraction
2.08M
Total gradient volume, incl. sample (mL)
11.6
Sample volume (mL)
0.3
Orientation
Bottom-up
Rotor type
SW 40 Ti
Speed (g)
270000
Duration (min)
960
Fraction volume (mL)
1
Fraction processing
None
Characterization: Protein analysis
Protein Concentration Method
Not determined
Western Blot
Antibody details provided?
Yes
Antibody dilution provided?
Yes
Lysis buffer provided?
Yes
Flow cytometry
Type of Flow cytometry
BD Influx/ Beckman Coulter CytoFLEX LX
Hardware adjustments
BD Influx: Adapted small particle detector
Calibration bead size
0.1 and 0.2
Antibody details provided?
No
Detected EV-associated proteins
CD9/ EGFR/ CD142
Characterization: Lipid analysis
No
Characterization: Particle analysis
Particle analysis: flow cytometry
Flow cytometer type
BD Influx/ Beckman Coulter Cytoflex LX
Hardware adjustment
BD Influx: adapted small particle detector
Calibration bead size
0.1 and 0.2
EV concentration
Yes
EV210279 2/2 Homo sapiens Blood plasma qEV column Libregts SFWM 2018 13%

Study summary

Full title
All authors
Libregts SFWM, Arkesteijn GJA, Németh A, Nolte-'t Hoen ENM, Wauben MHM
Journal
J Thromb Haemost
Abstract
Essentials Extracellular vesicles (EVs) in biological fluids are promising biomarkers for disease. F (show more...)Essentials Extracellular vesicles (EVs) in biological fluids are promising biomarkers for disease. Fluorescence-based flow cytometric analysis is suitable to detect low abundant EV subsets. Particles of non-interest can induce false-positive light scatter and fluorescent signals. Interference of particles of non-interest can be monitored by analyzing serial dilutions. (hide)
EV-METRIC
13% (32nd 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 data
Sample type
Blood plasma
Sample origin
Control condition
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
  • Gives a short, non-chronological overview of the
    different steps of the separation protocol.
    • dUC = (Differential) (ultra)centrifugation
    • DG = density gradient
    • UF = ultrafiltration
    • SEC = size-exclusion chromatography
    • IAF = immuno-affinity capture
Commercial method
Protein markers
EV: CD63/ CD9
non-EV: None
Proteomics
no
Show all info
Study aim
Technical analysis comparing/optimizing EV-related methods
Sample
Species
Homo sapiens
Sample Type
Blood plasma
Separation Method
Commercial kit
qEV column
Characterization: Protein analysis
Protein Concentration Method
Other/ Absorbance spectrometry (280 nm)
Western Blot
Antibody details provided?
Yes
Antibody dilution provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
CD9/ CD63
Characterization: Lipid analysis
No
Characterization: Particle analysis
Particle analysis: flow cytometry
Flow cytometer type
BD Influx/ Beckman Coulter Cytoflex LX
Hardware adjustment
BD Influx: adapted small particle detector
Calibration bead size
0.1 and 0.2
EV concentration
Yes
1 - 2 of 2
  • CM = Commercial method
  • dUC = differential ultracentrifugation
  • DG = density gradient
  • UF = ultrafiltration
  • SEC = size-exclusion chromatography
EV-TRACK ID
EV210279
species
Homo sapiens
sample type
Cell culture
Blood plasma
cell type
A431
NA
medium
EV-depleted medium
NA
condition
Control condition
Control condition
separation protocol
dUC/
Density gradient
qEV column
Exp. nr.
1
2
EV-METRIC %
63
13