Search > Results

You searched for: EV120001 (EV-TRACK ID)

Showing 1 - 1 of 1

Experiment number
  • If needed, multiple experiments were identified in a single publication based on differing sample types, isolation protocols and/or vesicle types of interest.
Species
  • Species of origin of the EVs.
Isolation protocol
  • Gives a short, non-chronological overview of the different steps of the isolation protocol.
    • dUC = differential ultracentrifugation
    • DG = density gradient
    • UF = ultrafiltration
    • SEC = size-exclusion chromatography
Details EV-TRACK ID Experiment nr. Species Sample type Isolation protocol First author Year EV-METRIC
EV120001 1/1 Oryctolagus cuniculus Cell culture supernatant 0.2 µm filter
Density cushion (valid.)
dUC
Iodixanol-DG (valid.)
Coleman BM 2012 78%

Study summary

Full title
All authors
Coleman BM, Hanssen E, Lawson VA, Hill AF
Journal
FASEB J
Abstract
Exosomes are small membrane-bound vesicles released from cells and found in vivo in most biological (show more...)Exosomes are small membrane-bound vesicles released from cells and found in vivo in most biological fluids. Functions reported for exosomes include cell-cell communication, roles in modulating immune responses, and roles in the transfer of pathogens such as prions. Here we investigated the molecular characteristics of the structure of exosomes that harbor prion infectivity to determine the native structure of exosomes and whether infected exosomes have a distinct structure. Cryo-electron tomography revealed the previously unidentified ultrastructural detail of exosomes with high resolution. Exosomes were found to be naturally spherical in shape and to have a diverse population that varies in size and internal structure, such as differences in the number of membrane structures. Exosomes isolated from prion-infected cells contained a significantly different population of exosomes with distinct structural features compared to control vesicles from mock-infected cells. Exosomes are highly structured vesicles that can modify their structure on altering their protein cargo. This finding provides further insight into the role that the exosomal protein cargo plays on influencing the structure of the vesicles as well as highlighting the diversity of exosomes and their relationship to biological processes. (hide)
EV-METRIC
78% (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
Isolation method: density gradient, at least as validation of results attributed to EVs
EV density
Isolation method: reporting of obtained EV density
ultracentrifugation specifics
Isolation 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
Focus vesicles
exosomes
Isolation protocol
Isolation protocol
  • Gives a short, non-chronological overview of the
    different steps of the isolation protocol.
    • dUC = differential ultracentrifugation
    • DG = density gradient
    • UF = ultrafiltration
    • SEC = size-exclusion chromatography
0.2 µm filter + Density cushion (valid.) + dUC + Iodixanol-DG (valid.)
Protein markers
EV: CD63/ Flotilin1/ TSG101
non-EV: Cell organelle protein/ Bcl2
Proteomics
no
TEM measurements
112+-12.7
Show all info
Study aim
Other/Ultrastructure
Sample
Species
Oryctolagus cuniculus
Sample Type
Cell culture supernatant
EV-harvesting Medium
EV Depleted
Isolation Method
Differential ultra centrifugation
Differential UC: filtering steps
Between 800 g and 10,000 g
Between 10,000 g and 50,000 g
Between 100,000 g and 150,000 g
Density gradient
Only used for validation of main results
1
Lowest density fraction
10
Highest density fraction
30
Orientation
Top-down
Speed (g)
10000
Filtration steps
0.22µm or 0.2µm
Characterization: Protein analysis
Western Blot
Detected EV-associated proteins
CD63/ Flotilin1/ TSG101
Detected contaminants
Cell organelle protein/ Bcl2
Characterization: Particle analysis
EM
EM-type
transmission EM/ cryo EM
Image type
Close-up, Wide-field
1 - 1 of 1