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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV100031	1/1	Homo sapiens	NAY	(d)(U)C Filtration IAF	Zumaquero E	2010	33%
Study summary Full title Exosomes from human lymphoblastoid B cells express enzymatically active CD38 that is associated with signaling complexes containing CD81, Hsc-70 and Lyn All authors Zumaquero E, Muñoz P, Cobo M, Lucena G, Pavón EJ, Martín A, Navarro P, García-Pérez A, Ariza-Veguillas A, Malavasi F, Sancho J, Zubiaur M Journal Exp Cell Res Abstract Exosome vesicles of endocytic origin are involved in communication between tumor and immune cells. I (show more...)Exosome vesicles of endocytic origin are involved in communication between tumor and immune cells. In addition, membrane rafts (MR) may support the sorting of proteins associated with exosomes. CD38 is found at the plasma membrane and in recycling endosomes, which are both redistributed toward the immunological synapse (IS) upon T cell antigen receptor (TCR) engagement. The data of this study provide evidence that CD38 is expressed on the surface of secreted exosomes derived from lymphoblastoid B cells. Exosomic CD38 is associated with the signaling molecules CD81, Hsc-70 and Lyn. Likewise, in MR, CD38 is associated with CD81, CD19, Lyn, Galphai-2, Hsc-70 and actin. Therefore, a high degree of overlap in the pattern of signaling proteins associated with CD38 in exosomes and MR exists. Exosomic and MR CD38, by virtue of these interactions, have signaling potential. Indeed, CD38 is enzymatically active in both exosomes and MR, and CD38 ligation induces Akt/PKB and Erk activation, which is accompanied by increased translocation of CD38 into MR. In conclusion, the present study indicates that CD38 localizes to MR, where it promotes cell signaling, and it is exported out of the cells through the exosome-mediated exocytic pathway, where it may act as an intercellular messenger. (hide) EV-METRIC 33% (75th 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 NAY Focus vesicles exosomes 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 (d)(U)C Filtration IAF Adj. k-factor 124.9 (pelleting) / 124.9 (washing) Protein markers EV: CD38/ Rab5/ Hsc70/ CD81/ Lyn/ MHC2 non-EV: Proteomics no Show all info Study aim Biogenesis/Sorting Sample Species Homo sapiens Sample Type Cell culture supernatant EV-harvesting Medium EV Depleted Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Below or equal to 800 g Between 800 g and 10,000 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: time(min) 180-720 Pelleting: rotor type AH650 Pelleting: adjusted k-factor 124.9 Wash: Rotor Type AH650 Wash: adjusted k-factor 124.9 Filtration steps 0.22µm or 0.2µm Immunoaffinity capture Selected surface protein(s) CD38, Lyn, CD81, Hsc70 Characterization: Protein analysis Western Blot Antibody details provided? No Detected EV-associated proteins CD81/ MHC2/ CD38/ Rab5/ Lyn/ Hsc70 ELISA Antibody details provided? No Detected EV-associated proteins MHC2/ CD38/ Rab5/ Lyn/ Hsc70 Characterization: Particle analysis EM EM-type immune EM EM protein CD38 Image type Close-up

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EV-TRACK ID	EV100031
species	Homo sapiens
sample type	Cell culture
cell type	NAY
condition	NAY
separation protocol	(d)(U)C Filtration IAF
Exp. nr.	1
EV-METRIC %	33