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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV230049	1/2	Homo sapiens	Brain gray matter	(d)(U)C DG Filtration	Muraoka S	2020	78%
Study summary Full title Proteomic and biological profiling of extracellular vesicles from Alzheimer's disease human brain tissues. All authors Muraoka S, DeLeo AM, Sethi MK, Yukawa-Takamatsu K, Yang Z, Ko J, Hogan JD, Ruan Z, You Y, Wang Y, Medalla M, Ikezu S, Chen M, Xia W, Gorantla S, Gendelman HE, Issadore D, Zaia J, Ikezu T Journal Alzheimers Dement Abstract Extracellular vesicles (EVs) from human Alzheimer's disease (AD) biospecimens contain amyloid beta ( (show more...)Extracellular vesicles (EVs) from human Alzheimer's disease (AD) biospecimens contain amyloid beta (Aβ) peptide and tau. While AD EVs are known to affect brain disease pathobiology, their biochemical and molecular characterizations remain ill defined. (hide) EV-METRIC 78% (50th 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 Brain gray matter 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 Filtration Protein markers EV: AB1-40/ AB1-42/ ANXA-5 non-EV: None Proteomics yes EV density (g/ml) 1.10-1.15 Show all info Study aim Function/Identification of content (omics approaches) Sample Species Homo sapiens Sample Type Brain gray matter Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Below or equal to 800 g Between 800 g and 10,000 g Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: rotor type SW 41 Ti Pelleting: speed (g) 100000 Density gradient Type Discontinuous Number of initial discontinuous layers 6 Lowest density fraction 0.475 M Highest density fraction 2.0 M Total gradient volume, incl. sample (mL) 14 Sample volume (mL) 2 Rotor type SW 41 Ti Speed (g) 200000 Duration (min) 1200 Fraction volume (mL) 2 Fraction processing Centrifugation Pelleting: volume per fraction 12 Pelleting: speed (g) 100000 Filtration steps Between 0.22 and 0.45 µm/ 0.2 or 0.22 µm Characterization: Protein analysis Protein Concentration Method BCA ELISA Detected EV-associated proteins AB1-40/ AB1-42/ ANXA-5 Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Modus Reported size (nm) 131 EV concentration Yes EM EM-type Transmission-EM Image type Close-up

	EV230049	2/2	Homo sapiens	Brain gray matter	(d)(U)C DG Filtration	Muraoka S	2020	78%
Study summary Full title Proteomic and biological profiling of extracellular vesicles from Alzheimer's disease human brain tissues. All authors Muraoka S, DeLeo AM, Sethi MK, Yukawa-Takamatsu K, Yang Z, Ko J, Hogan JD, Ruan Z, You Y, Wang Y, Medalla M, Ikezu S, Chen M, Xia W, Gorantla S, Gendelman HE, Issadore D, Zaia J, Ikezu T Journal Alzheimers Dement Abstract Extracellular vesicles (EVs) from human Alzheimer's disease (AD) biospecimens contain amyloid beta ( (show more...)Extracellular vesicles (EVs) from human Alzheimer's disease (AD) biospecimens contain amyloid beta (Aβ) peptide and tau. While AD EVs are known to affect brain disease pathobiology, their biochemical and molecular characterizations remain ill defined. (hide) EV-METRIC 78% (50th 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 Brain gray matter Sample origin Alzheimer's disease 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 Filtration Protein markers EV: AB1-40/ AB1-42/ ANXA5 non-EV: None Proteomics yes EV density (g/ml) 1.10-1.15 Show all info Study aim Function/Identification of content (omics approaches) Sample Species Homo sapiens Sample Type Brain gray matter Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Below or equal to 800 g Between 800 g and 10,000 g Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: rotor type SW 41 Ti Pelleting: speed (g) 100000 Density gradient Type Discontinuous Number of initial discontinuous layers 6 Lowest density fraction 0.475 M Highest density fraction 2.0 M Total gradient volume, incl. sample (mL) 14 Sample volume (mL) 2 Rotor type SW 41 Ti Speed (g) 200000 Duration (min) 1200 Fraction volume (mL) 2 Fraction processing Centrifugation Pelleting: volume per fraction 12 Pelleting: speed (g) 100000 Filtration steps Between 0.22 and 0.45 µm/ 0.2 or 0.22 µm Characterization: Protein analysis Protein Concentration Method BCA ELISA Detected EV-associated proteins AB1-40/ AB1-42/ ANXA5 Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Modus Reported size (nm) 122 EV concentration Yes EM EM-type Transmission-EM Image type Close-up

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EV-TRACK ID	EV230049
species	Homo sapiens
sample type	Brain gray matter
condition	Control condition	Alzheimer's disease
separation protocol	dUC/ Density gradient/ Filtration	dUC/ Density gradient/ Filtration
Exp. nr.	1	2
EV-METRIC %	78	78