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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV120085	1/1	Homo sapiens	Semen	(d)(U)C DG Filtration SEC	Ronquist GK	2012	44%
Study summary Full title Prostasomes are heterogeneous regarding size and appearance but affiliated to one DNA-containing exosome family All authors Ronquist GK, Larsson A, Stavreus-Evers A, Ronquist G Journal Prostate Abstract BACKGROUND: Prostate acinar epithelial cells release microvesicles (prostasomes) that possess pleiot (show more...)BACKGROUND: Prostate acinar epithelial cells release microvesicles (prostasomes) that possess pleiotropic biological effects relevant for successful fertilization. Prostasomes are formed in a similar way as exosomes but are heterogeneous as regards size and appearance. Like exosomes they are thought to be mediators of intercellular communication. METHODS: We prepared seminal prostasomes in accordance with the prevailing protocol for exosome preparation including passage through a 0.2 µm filter and centrifugation in a sucrose gradient. RESULTS: We compared the filterable prostasomes with those trapped on the filter (nonfilterable prostasomes) and, qualitatively, no conspicuous differences were apparent regarding ultrastructure and SDS-PAGE banding pattern. Moreover, both types of prostasomes contained DNA fragments and Western blot revealed presence of prostate specific membrane antigen (PSMA), CD38, and annexin A1. CONCLUSIONS: Reasonably, prostasomes could be included in the exosome family and be regarded as one entity containing chromosomal DNA. (hide) EV-METRIC 44% (82nd 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 Semen Sample origin NAY Focus vesicles exosomes / Prostasomes 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 DG Filtration SEC Adj. k-factor 126 (pelleting) Protein markers EV: Annexin1/ PSMA/ CD38 non-EV: Proteomics no EV density (g/ml) 1.13-1.26 Show all info Study aim Omics Sample Species Homo sapiens Sample Type Semen Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps 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: time(min) 120 Pelleting: rotor type 90Ti Pelleting: adjusted k-factor 126.0 Density gradient Lowest density fraction 0.25 Highest density fraction 2 Orientation Top-down Rotor type 90Ti Speed (g) 100000 Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Western Blot Antibody details provided? No Detected EV-associated proteins Annexin1/ PSMA/ CD38 ELISA Antibody details provided? No Detected EV-associated proteins Annexin1/ PSMA/ CD38 Characterization: Particle analysis EM EM-type transmission EM Image type Wide-field

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EV-TRACK ID	EV120085
species	Homo sapiens
sample type	Semen
condition	NAY
separation protocol	(d)(U)C DG Filtration SEC
Exp. nr.	1
EV-METRIC %	44