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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV230696	1/2	Bacillus subtilis	168	(d)(U)C Filtration UF	Kim Y	2016	29%
Study summary Full title Extracellular vesicle proteomes reflect developmental phases of Bacillus subtilis. All authors Kim Y, Edwards N, Fenselau C Journal Clin Proteomics Abstract Extracellular vesicles (EV) are spherical membrane-bound vesicles with nano-scale diameters, which a (show more...)Extracellular vesicles (EV) are spherical membrane-bound vesicles with nano-scale diameters, which are shed to the extracellular region by most eukaryotic and prokaryotic cells. Bacterial EV are proposed to contribute to intercellular communication, bacterial survival and human pathogenesis as a novel secretion system. EV have been characterized from many Gram-negative species and, more recently, from several vegetative Gram-positive bacteria. Further characterization of EV and their molecular cargos will contribute to understanding bacterial physiology and to developing therapeutic approaches. (hide) EV-METRIC 29% (67th 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 Vegetative 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 Filtration Ultrafiltration Protein markers EV: None non-EV: None Proteomics yes Show all info Study aim Biogenesis/cargo sorting Sample Species Bacillus subtilis Sample Type Cell culture supernatant EV-producing cells 168 EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Equal to or above 150,000 g Pelleting performed Yes Pelleting: rotor type 70Yi Pelleting: speed (g) 150000 Wash: time (min) 90 Wash: Rotor Type 70Yi Wash: speed (g) 150000 Filtration steps 0.22µm or 0.2µm Ultra filtration Cut-off size (kDa) 100 Membrane type NS Characterization: Protein analysis Protein Concentration Method BCA Proteomics database No Characterization: Lipid analysis Yes Characterization: Particle analysis EM EM-type Transmission-EM Image type Wide-field Report size (nm) 60

	EV230696	2/2	Bacillus subtilis	168	(d)(U)C Filtration UF	Kim Y	2016	29%
Study summary Full title Extracellular vesicle proteomes reflect developmental phases of Bacillus subtilis. All authors Kim Y, Edwards N, Fenselau C Journal Clin Proteomics Abstract Extracellular vesicles (EV) are spherical membrane-bound vesicles with nano-scale diameters, which a (show more...)Extracellular vesicles (EV) are spherical membrane-bound vesicles with nano-scale diameters, which are shed to the extracellular region by most eukaryotic and prokaryotic cells. Bacterial EV are proposed to contribute to intercellular communication, bacterial survival and human pathogenesis as a novel secretion system. EV have been characterized from many Gram-negative species and, more recently, from several vegetative Gram-positive bacteria. Further characterization of EV and their molecular cargos will contribute to understanding bacterial physiology and to developing therapeutic approaches. (hide) EV-METRIC 29% (67th 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 Sporulating 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 Filtration Ultrafiltration Protein markers EV: None non-EV: None Proteomics yes Show all info Study aim Biogenesis/cargo sorting Sample Species Bacillus subtilis Sample Type Cell culture supernatant EV-producing cells 168 EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Equal to or above 150,000 g Pelleting performed Yes Pelleting: rotor type 70Yi Pelleting: speed (g) 150000 Wash: time (min) 90 Wash: Rotor Type 70Yi Wash: speed (g) 150000 Filtration steps 0.22µm or 0.2µm Ultra filtration Cut-off size (kDa) 100 Membrane type NS Characterization: Protein analysis Protein Concentration Method BCA Proteomics database No Characterization: Lipid analysis Yes Characterization: Particle analysis EM EM-type Transmission-EM Image type Wide-field Report size (nm) 60

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EV-TRACK ID	EV230696
species	Bacillus subtilis
sample type	Cell culture
cell type	168
condition	Vegetative	Sporulating
separation protocol	dUC/ Filtration/ Ultrafiltration	dUC/ Filtration/ Ultrafiltration
Exp. nr.	1	2
EV-METRIC %	29	29