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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV210096	2/6	Homo sapiens	Urine	(d)(U)C Filtration	Royo, Felix	2016	38%
Study summary Full title Different EV enrichment methods suitable for clinical settings yield different subpopulations of urinary extracellular vesicles from human samples All authors Felix Royo, Patricia Zuñiga-Garcia, Pilar Sanchez-Mosquera, Ainara Egia, Amparo Perez, Ana Loizaga, Raquel Arceo, Isabel Lacasa, Ainara Rabade, Edurne Arrieta, Roberto Bilbao, Miguel Unda, Arkaitz Carracedo, Juan M Falcon-Perez Journal J Extracell Vesicles Abstract Urine sample analysis is irreplaceable as a non-invasive method for disease diagnosis and follow-up. (show more...)Urine sample analysis is irreplaceable as a non-invasive method for disease diagnosis and follow-up. However, in urine samples, non-degraded protein and RNA may be only found in urinary extracellular vesicles (uEVs). In recent years, various methods of uEV enrichment using low volumes of urine and unsophisticated equipment have been developed, with variable success. We compared the results of the differential ultracentrifugation procedure with 4 of these methods. The methods tested were a lectin-based purification, Exoquick (System Biosciences), Total Exosome Isolation from Invitrogen and an in-house modified procedure employing the Exosomal RNA Kit from Norgen Biotek Corp. The analysis of selected gene transcripts and protein markers of extracellular vesicles (EVs) revealed that each method isolates a different mixture of uEV protein markers. In our conditions, the extraction with Norgen's reagent achieved the best performance in terms of gene transcript and protein detection and reproducibility. By using this method, we were able to detect alterations of EVs protein markers in urine samples from prostate cancer adenoma patients. Taken together, our results show that the isolation of uEVs is feasible from small volumes of urine and avoiding ultracentrifugation, making easier the analysis in a clinical facility. However, caution should be taken in the selection of the enrichment method since they have a differential affinity for protein uEVs markers and by extension for different subpopulation of EVs. (hide) EV-METRIC 38% (73rd 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 Urine 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 (d)(U)C Filtration Protein markers EV: TSG101/ CD63/ Flotillin1/ CD9/ CD26/ CD10/ AQP2/ AIP non-EV: Tamm-Horsfall protein Proteomics no Show all info Study aim Technical analysis comparing/optimizing EV-related methods Sample Species Homo sapiens Sample Type Urine Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: time(min) 90 Pelleting: rotor type Type 70 Ti Pelleting: speed (g) 100000 Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method Not determined Western Blot Antibody details provided? No Detected EV-associated proteins Flotillin1/ CD9/ CD63/ CD26/ CD10/ AQP2/ AIP/ TSG101 Detected contaminants Tamm-Horsfall protein Characterization: RNA analysis RNA analysis Type (RT)(q)PCR;Capillary electrophoresis (e.g. Bioanalyzer) Database No Proteinase treatment No RNAse treatment No Characterization: Lipid analysis No EM EM-type Cryo-EM Image type Close-up, Wide-field

	EV210096	5/6	Homo sapiens	Urine	(d)(U)C Filtration Total Exosome Isolation	Royo, Felix	2016	38%
Study summary Full title Different EV enrichment methods suitable for clinical settings yield different subpopulations of urinary extracellular vesicles from human samples All authors Felix Royo, Patricia Zuñiga-Garcia, Pilar Sanchez-Mosquera, Ainara Egia, Amparo Perez, Ana Loizaga, Raquel Arceo, Isabel Lacasa, Ainara Rabade, Edurne Arrieta, Roberto Bilbao, Miguel Unda, Arkaitz Carracedo, Juan M Falcon-Perez Journal J Extracell Vesicles Abstract Urine sample analysis is irreplaceable as a non-invasive method for disease diagnosis and follow-up. (show more...)Urine sample analysis is irreplaceable as a non-invasive method for disease diagnosis and follow-up. However, in urine samples, non-degraded protein and RNA may be only found in urinary extracellular vesicles (uEVs). In recent years, various methods of uEV enrichment using low volumes of urine and unsophisticated equipment have been developed, with variable success. We compared the results of the differential ultracentrifugation procedure with 4 of these methods. The methods tested were a lectin-based purification, Exoquick (System Biosciences), Total Exosome Isolation from Invitrogen and an in-house modified procedure employing the Exosomal RNA Kit from Norgen Biotek Corp. The analysis of selected gene transcripts and protein markers of extracellular vesicles (EVs) revealed that each method isolates a different mixture of uEV protein markers. In our conditions, the extraction with Norgen's reagent achieved the best performance in terms of gene transcript and protein detection and reproducibility. By using this method, we were able to detect alterations of EVs protein markers in urine samples from prostate cancer adenoma patients. Taken together, our results show that the isolation of uEVs is feasible from small volumes of urine and avoiding ultracentrifugation, making easier the analysis in a clinical facility. However, caution should be taken in the selection of the enrichment method since they have a differential affinity for protein uEVs markers and by extension for different subpopulation of EVs. (hide) EV-METRIC 38% (73rd 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 Urine 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 (d)(U)C Filtration Commercial method Protein markers EV: TSG101/ CD63/ Flotillin1/ CD9/ CD26/ CD10/ AQP2/ AIP non-EV: Tamm-Horsfall protein Proteomics no Show all info Study aim Technical analysis comparing/optimizing EV-related methods Sample Species Homo sapiens Sample Type Urine Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Pelleting performed No Filtration steps 0.22µm or 0.2µm Commercial kit Total Exosome Isolation Characterization: Protein analysis Protein Concentration Method Not determined Western Blot Antibody details provided? No Detected EV-associated proteins CD26/ CD10/ AQP2/ AIP/ CD9/ CD63/ TSG101 Not detected EV-associated proteins Flotillin1 Detected contaminants Tamm-Horsfall protein Characterization: RNA analysis RNA analysis Type (RT)(q)PCR;Capillary electrophoresis (e.g. Bioanalyzer) Database No Proteinase treatment No RNAse treatment No Characterization: Lipid analysis No EM EM-type Cryo-EM Image type Close-up, Wide-field

	EV210096	6/6	Homo sapiens	Urine	(d)(U)C Filtration Extraction with biotinylated Solanum tuberosum (potato) lectin (STL) and streptavidin coated Dynabeads	Royo, Felix	2016	38%
Study summary Full title Different EV enrichment methods suitable for clinical settings yield different subpopulations of urinary extracellular vesicles from human samples All authors Felix Royo, Patricia Zuñiga-Garcia, Pilar Sanchez-Mosquera, Ainara Egia, Amparo Perez, Ana Loizaga, Raquel Arceo, Isabel Lacasa, Ainara Rabade, Edurne Arrieta, Roberto Bilbao, Miguel Unda, Arkaitz Carracedo, Juan M Falcon-Perez Journal J Extracell Vesicles Abstract Urine sample analysis is irreplaceable as a non-invasive method for disease diagnosis and follow-up. (show more...)Urine sample analysis is irreplaceable as a non-invasive method for disease diagnosis and follow-up. However, in urine samples, non-degraded protein and RNA may be only found in urinary extracellular vesicles (uEVs). In recent years, various methods of uEV enrichment using low volumes of urine and unsophisticated equipment have been developed, with variable success. We compared the results of the differential ultracentrifugation procedure with 4 of these methods. The methods tested were a lectin-based purification, Exoquick (System Biosciences), Total Exosome Isolation from Invitrogen and an in-house modified procedure employing the Exosomal RNA Kit from Norgen Biotek Corp. The analysis of selected gene transcripts and protein markers of extracellular vesicles (EVs) revealed that each method isolates a different mixture of uEV protein markers. In our conditions, the extraction with Norgen's reagent achieved the best performance in terms of gene transcript and protein detection and reproducibility. By using this method, we were able to detect alterations of EVs protein markers in urine samples from prostate cancer adenoma patients. Taken together, our results show that the isolation of uEVs is feasible from small volumes of urine and avoiding ultracentrifugation, making easier the analysis in a clinical facility. However, caution should be taken in the selection of the enrichment method since they have a differential affinity for protein uEVs markers and by extension for different subpopulation of EVs. (hide) EV-METRIC 38% (73rd 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 Urine 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 (d)(U)C Filtration Extraction with biotinylated Solanum tuberosum (potato) lectin (STL) and streptavidin coated Dynabeads Protein markers EV: TSG101/ CD63/ AIP/ CD26/ CD10/ AQP2/ Flotillin1/ CD9 non-EV: Tamm-Horsfall protein Proteomics no Show all info Study aim Technical analysis comparing/optimizing EV-related methods Sample Species Homo sapiens Sample Type Urine Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Pelleting performed No Filtration steps 0.22µm or 0.2µm Other Name other separation method Extraction with biotinylated Solanum tuberosum (potato) lectin (STL) and streptavidin coated Dynabeads Characterization: Protein analysis Protein Concentration Method Not determined Western Blot Antibody details provided? No Detected EV-associated proteins Flotillin1/ CD26/ CD10/ AQP2/ TSG101/ CD9/ CD63 Not detected EV-associated proteins AIP Detected contaminants Tamm-Horsfall protein Characterization: RNA analysis RNA analysis Type (RT)(q)PCR;Capillary electrophoresis (e.g. Bioanalyzer) Database No Proteinase treatment No RNAse treatment No Characterization: Lipid analysis No EM EM-type Cryo-EM Image type Close-up, Wide-field

	EV210096	1/6	Homo sapiens	Urine	(d)(U)C Filtration Urine Exosome RNA Isolation Kit (Norgen Biotek Corp)	Royo, Felix	2016	25%
Study summary Full title Different EV enrichment methods suitable for clinical settings yield different subpopulations of urinary extracellular vesicles from human samples All authors Felix Royo, Patricia Zuñiga-Garcia, Pilar Sanchez-Mosquera, Ainara Egia, Amparo Perez, Ana Loizaga, Raquel Arceo, Isabel Lacasa, Ainara Rabade, Edurne Arrieta, Roberto Bilbao, Miguel Unda, Arkaitz Carracedo, Juan M Falcon-Perez Journal J Extracell Vesicles Abstract Urine sample analysis is irreplaceable as a non-invasive method for disease diagnosis and follow-up. (show more...)Urine sample analysis is irreplaceable as a non-invasive method for disease diagnosis and follow-up. However, in urine samples, non-degraded protein and RNA may be only found in urinary extracellular vesicles (uEVs). In recent years, various methods of uEV enrichment using low volumes of urine and unsophisticated equipment have been developed, with variable success. We compared the results of the differential ultracentrifugation procedure with 4 of these methods. The methods tested were a lectin-based purification, Exoquick (System Biosciences), Total Exosome Isolation from Invitrogen and an in-house modified procedure employing the Exosomal RNA Kit from Norgen Biotek Corp. The analysis of selected gene transcripts and protein markers of extracellular vesicles (EVs) revealed that each method isolates a different mixture of uEV protein markers. In our conditions, the extraction with Norgen's reagent achieved the best performance in terms of gene transcript and protein detection and reproducibility. By using this method, we were able to detect alterations of EVs protein markers in urine samples from prostate cancer adenoma patients. Taken together, our results show that the isolation of uEVs is feasible from small volumes of urine and avoiding ultracentrifugation, making easier the analysis in a clinical facility. However, caution should be taken in the selection of the enrichment method since they have a differential affinity for protein uEVs markers and by extension for different subpopulation of EVs. (hide) EV-METRIC 25% (56th 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 Urine Sample origin Prostate cancer 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 (d)(U)C Filtration Commercial method Protein markers EV: TSG101/ CD63/ Flotillin1/ CD9/ CD26/ CD10/ AQP2/ AIP non-EV: Tamm-Horsfall protein Proteomics no Show all info Study aim Technical analysis comparing/optimizing EV-related methods Sample Species Homo sapiens Sample Type Urine Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Pelleting performed No Filtration steps 0.22µm or 0.2µm Commercial kit Urine Exosome RNA Isolation Kit (Norgen Biotek Corp) Characterization: Protein analysis Protein Concentration Method Not determined Western Blot Antibody details provided? No Detected EV-associated proteins Flotillin1/ CD9/ CD63/ CD26/ CD10/ AQP2/ AIP/ TSG101 Detected contaminants Tamm-Horsfall protein Characterization: RNA analysis RNA analysis Type (RT)(q)PCR Database No Proteinase treatment No RNAse treatment No Characterization: Lipid analysis No

	EV210096	3/6	Homo sapiens	Urine	(d)(U)C Filtration ExoQuick	Royo, Felix	2016	25%
Study summary Full title Different EV enrichment methods suitable for clinical settings yield different subpopulations of urinary extracellular vesicles from human samples All authors Felix Royo, Patricia Zuñiga-Garcia, Pilar Sanchez-Mosquera, Ainara Egia, Amparo Perez, Ana Loizaga, Raquel Arceo, Isabel Lacasa, Ainara Rabade, Edurne Arrieta, Roberto Bilbao, Miguel Unda, Arkaitz Carracedo, Juan M Falcon-Perez Journal J Extracell Vesicles Abstract Urine sample analysis is irreplaceable as a non-invasive method for disease diagnosis and follow-up. (show more...)Urine sample analysis is irreplaceable as a non-invasive method for disease diagnosis and follow-up. However, in urine samples, non-degraded protein and RNA may be only found in urinary extracellular vesicles (uEVs). In recent years, various methods of uEV enrichment using low volumes of urine and unsophisticated equipment have been developed, with variable success. We compared the results of the differential ultracentrifugation procedure with 4 of these methods. The methods tested were a lectin-based purification, Exoquick (System Biosciences), Total Exosome Isolation from Invitrogen and an in-house modified procedure employing the Exosomal RNA Kit from Norgen Biotek Corp. The analysis of selected gene transcripts and protein markers of extracellular vesicles (EVs) revealed that each method isolates a different mixture of uEV protein markers. In our conditions, the extraction with Norgen's reagent achieved the best performance in terms of gene transcript and protein detection and reproducibility. By using this method, we were able to detect alterations of EVs protein markers in urine samples from prostate cancer adenoma patients. Taken together, our results show that the isolation of uEVs is feasible from small volumes of urine and avoiding ultracentrifugation, making easier the analysis in a clinical facility. However, caution should be taken in the selection of the enrichment method since they have a differential affinity for protein uEVs markers and by extension for different subpopulation of EVs. (hide) EV-METRIC 25% (56th 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 Urine 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 (d)(U)C Filtration Commercial method Protein markers EV: TSG101/ CD63/ CD10/ AQP2/ AIP/ CD26/ Flotillin1/ CD9 non-EV: Tamm-Horsfall protein Proteomics no Show all info Study aim Technical analysis comparing/optimizing EV-related methods Sample Species Homo sapiens Sample Type Urine Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Pelleting performed No Filtration steps 0.22µm or 0.2µm Commercial kit ExoQuick Characterization: Protein analysis Protein Concentration Method Not determined Western Blot Antibody details provided? No Detected EV-associated proteins CD9/ CD63/ CD26 Not detected EV-associated proteins Flotillin1/ CD10/ AQP2/ AIP/ TSG101 Detected contaminants Tamm-Horsfall protein Characterization: RNA analysis RNA analysis Type (RT)(q)PCR;Capillary electrophoresis (e.g. Bioanalyzer) Database No Proteinase treatment No RNAse treatment No Characterization: Lipid analysis No EM EM-type Cryo-EM Image type Wide-field

	EV210096	4/6	Homo sapiens	Urine	(d)(U)C Filtration Urine Exosome RNA Isolation Kit (Norgen Biotek Corp)	Royo, Felix	2016	25%
Study summary Full title Different EV enrichment methods suitable for clinical settings yield different subpopulations of urinary extracellular vesicles from human samples All authors Felix Royo, Patricia Zuñiga-Garcia, Pilar Sanchez-Mosquera, Ainara Egia, Amparo Perez, Ana Loizaga, Raquel Arceo, Isabel Lacasa, Ainara Rabade, Edurne Arrieta, Roberto Bilbao, Miguel Unda, Arkaitz Carracedo, Juan M Falcon-Perez Journal J Extracell Vesicles Abstract Urine sample analysis is irreplaceable as a non-invasive method for disease diagnosis and follow-up. (show more...)Urine sample analysis is irreplaceable as a non-invasive method for disease diagnosis and follow-up. However, in urine samples, non-degraded protein and RNA may be only found in urinary extracellular vesicles (uEVs). In recent years, various methods of uEV enrichment using low volumes of urine and unsophisticated equipment have been developed, with variable success. We compared the results of the differential ultracentrifugation procedure with 4 of these methods. The methods tested were a lectin-based purification, Exoquick (System Biosciences), Total Exosome Isolation from Invitrogen and an in-house modified procedure employing the Exosomal RNA Kit from Norgen Biotek Corp. The analysis of selected gene transcripts and protein markers of extracellular vesicles (EVs) revealed that each method isolates a different mixture of uEV protein markers. In our conditions, the extraction with Norgen's reagent achieved the best performance in terms of gene transcript and protein detection and reproducibility. By using this method, we were able to detect alterations of EVs protein markers in urine samples from prostate cancer adenoma patients. Taken together, our results show that the isolation of uEVs is feasible from small volumes of urine and avoiding ultracentrifugation, making easier the analysis in a clinical facility. However, caution should be taken in the selection of the enrichment method since they have a differential affinity for protein uEVs markers and by extension for different subpopulation of EVs. (hide) EV-METRIC 25% (56th 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 Urine 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 (d)(U)C Filtration Commercial method Protein markers EV: TSG101/ CD63/ Flotillin1/ CD9/ CD26/ CD10/ AQP2/ AIP non-EV: Tamm-Horsfall protein Proteomics no Show all info Study aim Technical analysis comparing/optimizing EV-related methods Sample Species Homo sapiens Sample Type Urine Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Pelleting performed No Filtration steps 0.22µm or 0.2µm Commercial kit Urine Exosome RNA Isolation Kit (Norgen Biotek Corp) Characterization: Protein analysis Protein Concentration Method Not determined Western Blot Antibody details provided? No Detected EV-associated proteins Flotillin1/ CD26/ CD10/ AQP2/ AIP/ CD9/ CD63/ TSG101 Detected contaminants Tamm-Horsfall protein Characterization: RNA analysis RNA analysis Type (RT)(q)PCR;Capillary electrophoresis (e.g. Bioanalyzer) Database No Proteinase treatment No RNAse treatment No Characterization: Lipid analysis No EM EM-type Cryo-EM Image type Wide-field

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EV-TRACK ID	EV210096
species	Homo sapiens
sample type	Urine
condition	Control condition	Control condition	Control condition	Prostate cancer	Control condition	Control condition
separation protocol	(d)(U)C Filtration	(d)(U)C Filtration Total Exosome Isolation	(d)(U)C Filtration Extraction with biotinylated Solanum tuberosum (potato) lectin (STL) and streptavidin coated Dynabeads	(d)(U)C Filtration Urine Exosome RNA Isolation Kit (Norgen Biotek Corp)	(d)(U)C Filtration ExoQuick	(d)(U)C Filtration Urine Exosome RNA Isolation Kit (Norgen Biotek Corp)
Exp. nr.	2	5	6	1	3	4
EV-METRIC %	38	38	38	25	25	25