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You searched for: EV170046 (EV-TRACK ID)
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Details | EV-TRACK ID | Experiment nr. | Species | Sample type | separation protocol | First author | Year | EV-METRIC |
---|---|---|---|---|---|---|---|---|
EV170046 | 1/2 | Sus scrofa | Cell culture supernatant |
DG (d)(U)C UF |
Klingeborn M | 2017 | 88% | |
Study summaryFull title
All authors
Klingeborn M, Dismuke WM, Skiba NP, Kelly U, Stamer WD, Bowes Rickman C
Journal
Sci Rep
Abstract
The retinal pigmented epithelium (RPE) forms the outer blood-retinal barrier in the eye and its pola (show more...)
EV-METRIC
88% (98th 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
Primary retinal pigmented epithelium cells
Sample origin
Basolateral side of polarized layer
Focus vesicles
exosome
Separation protocol
Separation protocol
DG
(d)(U)C UF Adj. k-factor
253.9 (pelleting) / 253.9 (washing)
Protein markers
EV: TSG101/ Syntenin-1
non-EV: Calreticulin Proteomics
yes
Show all info
Study aim
New methodological development, Identification of content (omics approaches)
Sample
Species
Sus scrofa
Sample Type
Cell culture supernatant
Sample Condition
Basolateral side of polarized layer
EV-producing cells
Primary retinal pigmented epithelium cells
EV-harvesting Medium
EV-depleted serum
Preparation of EDS
overnight (16h) at >=100,000g
Separation Method
Differential ultracentrifugation
centrifugation steps
Between 800 g and 10,000 g
Between 100,000 g and 150,000 g Obtain an EV pellet :
Yes
Pelleting: time(min)
90
Pelleting: rotor type
SW 28
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
253.9
Wash: time (min)
90
Wash: Rotor Type
SW 28
Wash: speed (g)
100000
Wash: adjusted k-factor
253.9
Density gradient
Only used for validation of main results
Yes
Density medium
Iodixanol
Type
Discontinuous
Number of initial discontinuous layers
4
Lowest density fraction
0.05
Highest density fraction
0.4
Sample volume (mL)
1
Orientation
Bottom-up (sample migrates upwards)
Rotor type
SW 41 Ti
Speed (g)
100000
Duration (min)
1080
Fraction volume (mL)
1
Fraction processing
Centrifugation
Pelleting: volume per fraction
12
Pelleting: duration (min)
90
Pelleting: rotor type
SW 41 Ti
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
255.8
Ultra filtration
Cut-off size (kDa)
100
Membrane type
Polyethersulfone (PES)
Characterization: Protein analysis
Protein Concentration Method
Pierce 660nm Protein Assay
Western Blot
Antibody details provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
TSG101, Syntenin-1
Not detected contaminants
Calreticulin
Proteomics
Proteomics database
No
Characterization: Particle analysis
NTA
Report type
Modus
Reported size (nm)
132.2±13.1
EM
EM-type
Transmission-EM
Image type
Wide-field
|
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EV170046 | 2/2 | Sus scrofa | Cell culture supernatant |
DG (d)(U)C UF |
Klingeborn M | 2017 | 88% | |
Study summaryFull title
All authors
Klingeborn M, Dismuke WM, Skiba NP, Kelly U, Stamer WD, Bowes Rickman C
Journal
Sci Rep
Abstract
The retinal pigmented epithelium (RPE) forms the outer blood-retinal barrier in the eye and its pola (show more...)
EV-METRIC
88% (98th 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
Primary retinal pigmented epithelium cells
Sample origin
Apical side of polarized layer
Focus vesicles
exosome
Separation protocol
Separation protocol
DG
(d)(U)C UF Adj. k-factor
253.9 (pelleting) / 253.9 (washing)
Protein markers
EV: TSG101/ Syntenin-1
non-EV: Calreticulin Proteomics
yes
Show all info
Study aim
New methodological development, Identification of content (omics approaches)
Sample
Species
Sus scrofa
Sample Type
Cell culture supernatant
Sample Condition
Apical side of polarized layer
EV-producing cells
Primary retinal pigmented epithelium cells
EV-harvesting Medium
EV-depleted serum
Preparation of EDS
overnight (16h) at >=100,000g
Separation Method
Differential ultracentrifugation
centrifugation steps
Between 800 g and 10,000 g
Between 100,000 g and 150,000 g Obtain an EV pellet :
Yes
Pelleting: time(min)
90
Pelleting: rotor type
SW 28
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
253.9
Wash: time (min)
90
Wash: Rotor Type
SW 28
Wash: speed (g)
100000
Wash: adjusted k-factor
253.9
Density gradient
Only used for validation of main results
Yes
Density medium
Iodixanol
Type
Discontinuous
Number of initial discontinuous layers
4
Lowest density fraction
0.05
Highest density fraction
0.4
Sample volume (mL)
1
Orientation
Bottom-up (sample migrates upwards)
Rotor type
SW 41 Ti
Speed (g)
100000
Duration (min)
1080
Fraction volume (mL)
1
Fraction processing
Centrifugation
Pelleting: volume per fraction
12
Pelleting: duration (min)
90
Pelleting: rotor type
SW 41 Ti
Pelleting: speed (g)
100000
Pelleting: adjusted k-factor
255.8
Ultra filtration
Cut-off size (kDa)
100
Membrane type
Polyethersulfone (PES)
Characterization: Protein analysis
Protein Concentration Method
Pierce 660nm Protein Assay
Western Blot
Antibody details provided?
Yes
Lysis buffer provided?
Yes
Detected EV-associated proteins
TSG101, Syntenin-1
Not detected contaminants
Calreticulin
Proteomics
Proteomics database
No
Characterization: Particle analysis
NTA
Report type
Modus
Reported size (nm)
118.6±9.9
EM
EM-type
Transmission-EM
Image type
Close-up
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