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You searched for: EV150104 (EV-TRACK ID)

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Experiment number
  • If needed, multiple experiments were identified in a single publication based on differing sample types, isolation protocols and/or vesicle types of interest.
Species
  • Species of origin of the EVs.
Isolation protocol
  • Gives a short, non-chronological overview of the different steps of the isolation protocol.
    • dUC = differential ultracentrifugation
    • DG = density gradient
    • UF = ultrafiltration
    • SEC = size-exclusion chromatography
Details EV-TRACK ID Experiment nr. Species Sample type Isolation protocol First author Year EV-METRIC
EV150104 1/4 Homo sapiens Urine (Differential) (ultra)centrifugation
Density cushion
Pocsfalvi G 2015 44%

Study summary

Full title
All authors
Pocsfalvi G, Raj DA, Fiume I, Vilasi A, Trepiccione F, Capasso G.
Journal
Proteomics Clin Appl
Abstract
PURPOSE: Recent findings indicate that urinary extracellular vesicles (EVs) might reflect the pathop (show more...)PURPOSE: Recent findings indicate that urinary extracellular vesicles (EVs) might reflect the pathophysiological state of urinary system; and that EVs-induced ciliary signaling is a possible mechanism of intercellular communication within the tract. Here, we aimed to analyze the protein expression of urinary EVs during autosomal dominant polycystic kidney disease (ADPKD). EXPERIMENTAL DESIGN: EVs were isolated from pooled urine samples of healthy control and ADPKD patients at two different stages of the disease and under tolvaptan treatment using the double-cushion ultracentrifugation method. Proteins were identified and quantified by iTRAQ and multidimensional protein identification technology (MudPIT)-based quantitative proteomics. RESULTS: Quantitative analyses identified 83 differentially expressed EV proteins. Many of these have apical membrane origin and are involved in signal transduction pathways of primary cilia, Ca(2+) -activated signaling, cell-cycle regulation, and cell differentiation. CONCLUSIONS AND CLINICAL RELEVANCE: The reduced AQP-2 and the increased APO-A1 levels observed in all ADPKD-affected groups may reflects the impaired renal concentrating capability of these patients and correlated with estimated glomerular filtration rate decline. The levels of some upregulated proteins involved in Ca(2+) -activated signaling declined upon tolvaptan treatment. The results obtained suggest that the quantitative proteomics of urinary EVs might be useful to monitor proteins difficult to access noninvasively, and thus advance our understanding of urinary tract physiology and pathology. (hide)
EV-METRIC
44% (85th 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
electron microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density gradient
Isolation method: density gradient, at least as validation of results attributed to EVs
EV density
Isolation method: reporting of obtained EV density
ultracentrifugation specifics
Isolation 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
Isolation protocol
Isolation protocol
  • Gives a short, non-chronological overview of the
    different steps of the isolation protocol.
    • dUC = differential ultracentrifugation
    • DG = density gradient
    • UF = ultrafiltration
    • SEC = size-exclusion chromatography
(Differential) (ultra)centrifugation + Density cushion
Protein markers
EV: None
non-EV: None
Proteomics
yes
Show all info
Study aim
Biomarker
Sample
Species
Homo sapiens
Sample Type
Urine
Sample Condition
Control condition
Isolation Method
Differential ultracentrifugation
dUC: centrifugation steps
Below or equal to 800 g
Between 800 g and 10,000 g
Between 10,000 g and 50,000 g
Equal to or above 150,000 g
Pelleting: time(min)
60
Pelleting: rotor type
Not specified
Pelleting: speed (g)
200000
Density cushion
Density medium
Sucrose
Characterization: Protein analysis
Protein Concentration Method
microBCA
Western Blot
Detected EV-associated proteins
CD9/ NHE3/ AQP2/ PKD11/ PKD22/ TSG101/ Alix
Detected contaminants
Not detected contaminants
Albumin/ Uromodulin
Proteomics database
No
Characterization: Particle analysis
NTA
Report type
Modus
Reported size (nm)
95
EM
EM-type
Transmission-EM
Image type
Wide-field
EV150104 2/4 Homo sapiens Urine (Differential) (ultra)centrifugation
Density cushion
Pocsfalvi G 2015 33%

Study summary

Full title
All authors
Pocsfalvi G, Raj DA, Fiume I, Vilasi A, Trepiccione F, Capasso G.
Journal
Proteomics Clin Appl
Abstract
PURPOSE: Recent findings indicate that urinary extracellular vesicles (EVs) might reflect the pathop (show more...)PURPOSE: Recent findings indicate that urinary extracellular vesicles (EVs) might reflect the pathophysiological state of urinary system; and that EVs-induced ciliary signaling is a possible mechanism of intercellular communication within the tract. Here, we aimed to analyze the protein expression of urinary EVs during autosomal dominant polycystic kidney disease (ADPKD). EXPERIMENTAL DESIGN: EVs were isolated from pooled urine samples of healthy control and ADPKD patients at two different stages of the disease and under tolvaptan treatment using the double-cushion ultracentrifugation method. Proteins were identified and quantified by iTRAQ and multidimensional protein identification technology (MudPIT)-based quantitative proteomics. RESULTS: Quantitative analyses identified 83 differentially expressed EV proteins. Many of these have apical membrane origin and are involved in signal transduction pathways of primary cilia, Ca(2+) -activated signaling, cell-cycle regulation, and cell differentiation. CONCLUSIONS AND CLINICAL RELEVANCE: The reduced AQP-2 and the increased APO-A1 levels observed in all ADPKD-affected groups may reflects the impaired renal concentrating capability of these patients and correlated with estimated glomerular filtration rate decline. The levels of some upregulated proteins involved in Ca(2+) -activated signaling declined upon tolvaptan treatment. The results obtained suggest that the quantitative proteomics of urinary EVs might be useful to monitor proteins difficult to access noninvasively, and thus advance our understanding of urinary tract physiology and pathology. (hide)
EV-METRIC
33% (66th 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
electron microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density gradient
Isolation method: density gradient, at least as validation of results attributed to EVs
EV density
Isolation method: reporting of obtained EV density
ultracentrifugation specifics
Isolation 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
autosomal dominant polycystic kidney disease (late stage)
Focus vesicles
extracellular vesicle
Isolation protocol
Isolation protocol
  • Gives a short, non-chronological overview of the
    different steps of the isolation protocol.
    • dUC = differential ultracentrifugation
    • DG = density gradient
    • UF = ultrafiltration
    • SEC = size-exclusion chromatography
(Differential) (ultra)centrifugation + Density cushion
Protein markers
EV: None
non-EV: None
Proteomics
yes
EV density (g/ml)
Show all info
Study aim
Biomarker
Sample
Species
Homo sapiens
Sample Type
Urine
Sample Condition
autosomal dominant polycystic kidney disease (late stage)
Isolation Method
Differential ultracentrifugation
dUC: centrifugation steps
Below or equal to 800 g
Between 800 g and 10,000 g
Between 10,000 g and 50,000 g
Equal to or above 150,000 g
Pelleting: time(min)
60
Pelleting: rotor type
Not specified
Pelleting: speed (g)
200000
Density gradient
Only used for validation of main results
Density medium
Type
Number of initial discontinuous layers
Lowest density fraction
Highest density fraction
Total gradient volume, incl. sample (mL)
Sample volume (mL)
Orientation
Rotor type
Speed (g)
Duration (min)
Fraction volume (mL)
Fraction processing
Pelleting: volume per fraction
Pelleting: duration (min)
Pelleting: rotor type
Pelleting: speed (g)
Pelleting-wash: volume per pellet (mL)
Pelleting-wash: duration (min)
Pelleting-wash: speed (g)
Density cushion
Density medium
Sucrose
Characterization: Protein analysis
Protein Concentration Method
microBCA
Western Blot
Detected EV-associated proteins
Alix/ NHE3/ AQP2/ CD9/ TSG101
Not detected EV-associated proteins
PKD1/ PKD2
Detected contaminants
Not detected contaminants
Albumin/ Uromodulin
Proteomics database
No
Characterization: Particle analysis
EM
EM-type
EV concentration
EV150104 3/4 Homo sapiens Urine (Differential) (ultra)centrifugation
Density cushion
Pocsfalvi G 2015 33%

Study summary

Full title
All authors
Pocsfalvi G, Raj DA, Fiume I, Vilasi A, Trepiccione F, Capasso G.
Journal
Proteomics Clin Appl
Abstract
PURPOSE: Recent findings indicate that urinary extracellular vesicles (EVs) might reflect the pathop (show more...)PURPOSE: Recent findings indicate that urinary extracellular vesicles (EVs) might reflect the pathophysiological state of urinary system; and that EVs-induced ciliary signaling is a possible mechanism of intercellular communication within the tract. Here, we aimed to analyze the protein expression of urinary EVs during autosomal dominant polycystic kidney disease (ADPKD). EXPERIMENTAL DESIGN: EVs were isolated from pooled urine samples of healthy control and ADPKD patients at two different stages of the disease and under tolvaptan treatment using the double-cushion ultracentrifugation method. Proteins were identified and quantified by iTRAQ and multidimensional protein identification technology (MudPIT)-based quantitative proteomics. RESULTS: Quantitative analyses identified 83 differentially expressed EV proteins. Many of these have apical membrane origin and are involved in signal transduction pathways of primary cilia, Ca(2+) -activated signaling, cell-cycle regulation, and cell differentiation. CONCLUSIONS AND CLINICAL RELEVANCE: The reduced AQP-2 and the increased APO-A1 levels observed in all ADPKD-affected groups may reflects the impaired renal concentrating capability of these patients and correlated with estimated glomerular filtration rate decline. The levels of some upregulated proteins involved in Ca(2+) -activated signaling declined upon tolvaptan treatment. The results obtained suggest that the quantitative proteomics of urinary EVs might be useful to monitor proteins difficult to access noninvasively, and thus advance our understanding of urinary tract physiology and pathology. (hide)
EV-METRIC
33% (66th 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
electron microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density gradient
Isolation method: density gradient, at least as validation of results attributed to EVs
EV density
Isolation method: reporting of obtained EV density
ultracentrifugation specifics
Isolation 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
autosomal dominant polycystic kidney disease (early stage)
Focus vesicles
extracellular vesicle
Isolation protocol
Isolation protocol
  • Gives a short, non-chronological overview of the
    different steps of the isolation protocol.
    • dUC = differential ultracentrifugation
    • DG = density gradient
    • UF = ultrafiltration
    • SEC = size-exclusion chromatography
(Differential) (ultra)centrifugation + Density cushion
Protein markers
EV: None
non-EV: None
Proteomics
yes
EV density (g/ml)
Show all info
Study aim
Biomarker
Sample
Species
Homo sapiens
Sample Type
Urine
Sample Condition
autosomal dominant polycystic kidney disease (early stage)
Isolation Method
Differential ultracentrifugation
dUC: centrifugation steps
Below or equal to 800 g
Between 800 g and 10,000 g
Between 10,000 g and 50,000 g
Equal to or above 150,000 g
Pelleting: time(min)
60
Pelleting: rotor type
Not specified
Pelleting: speed (g)
200000
Density gradient
Only used for validation of main results
Density medium
Type
Number of initial discontinuous layers
Lowest density fraction
Highest density fraction
Total gradient volume, incl. sample (mL)
Sample volume (mL)
Orientation
Rotor type
Speed (g)
Duration (min)
Fraction volume (mL)
Fraction processing
Pelleting: volume per fraction
Pelleting: duration (min)
Pelleting: rotor type
Pelleting: speed (g)
Pelleting-wash: volume per pellet (mL)
Pelleting-wash: duration (min)
Pelleting-wash: speed (g)
Density cushion
Density medium
Sucrose
Characterization: Protein analysis
Protein Concentration Method
microBCA
Western Blot
Detected EV-associated proteins
Alix/ NHE3/ AQP2/ CD9/ TSG101
Not detected EV-associated proteins
PKD1/ PKD2
Detected contaminants
Not detected contaminants
Albumin/ Uromodulin
Proteomics database
No
Characterization: Particle analysis
EM
EM-type
EV concentration
EV150104 4/4 Homo sapiens Urine (Differential) (ultra)centrifugation
Density cushion
Pocsfalvi G 2015 33%

Study summary

Full title
All authors
Pocsfalvi G, Raj DA, Fiume I, Vilasi A, Trepiccione F, Capasso G.
Journal
Proteomics Clin Appl
Abstract
PURPOSE: Recent findings indicate that urinary extracellular vesicles (EVs) might reflect the pathop (show more...)PURPOSE: Recent findings indicate that urinary extracellular vesicles (EVs) might reflect the pathophysiological state of urinary system; and that EVs-induced ciliary signaling is a possible mechanism of intercellular communication within the tract. Here, we aimed to analyze the protein expression of urinary EVs during autosomal dominant polycystic kidney disease (ADPKD). EXPERIMENTAL DESIGN: EVs were isolated from pooled urine samples of healthy control and ADPKD patients at two different stages of the disease and under tolvaptan treatment using the double-cushion ultracentrifugation method. Proteins were identified and quantified by iTRAQ and multidimensional protein identification technology (MudPIT)-based quantitative proteomics. RESULTS: Quantitative analyses identified 83 differentially expressed EV proteins. Many of these have apical membrane origin and are involved in signal transduction pathways of primary cilia, Ca(2+) -activated signaling, cell-cycle regulation, and cell differentiation. CONCLUSIONS AND CLINICAL RELEVANCE: The reduced AQP-2 and the increased APO-A1 levels observed in all ADPKD-affected groups may reflects the impaired renal concentrating capability of these patients and correlated with estimated glomerular filtration rate decline. The levels of some upregulated proteins involved in Ca(2+) -activated signaling declined upon tolvaptan treatment. The results obtained suggest that the quantitative proteomics of urinary EVs might be useful to monitor proteins difficult to access noninvasively, and thus advance our understanding of urinary tract physiology and pathology. (hide)
EV-METRIC
33% (66th 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
electron microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density gradient
Isolation method: density gradient, at least as validation of results attributed to EVs
EV density
Isolation method: reporting of obtained EV density
ultracentrifugation specifics
Isolation 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
autosomal dominant polycystic kidney disease (tolvaptan treatment)
Focus vesicles
extracellular vesicle
Isolation protocol
Isolation protocol
  • Gives a short, non-chronological overview of the
    different steps of the isolation protocol.
    • dUC = differential ultracentrifugation
    • DG = density gradient
    • UF = ultrafiltration
    • SEC = size-exclusion chromatography
(Differential) (ultra)centrifugation + Density cushion
Protein markers
EV: None
non-EV: None
Proteomics
yes
EV density (g/ml)
Show all info
Study aim
Biomarker
Sample
Species
Homo sapiens
Sample Type
Urine
Sample Condition
autosomal dominant polycystic kidney disease (tolvaptan treatment)
Isolation Method
Differential ultracentrifugation
dUC: centrifugation steps
Below or equal to 800 g
Between 800 g and 10,000 g
Between 10,000 g and 50,000 g
Equal to or above 150,000 g
Pelleting: time(min)
60
Pelleting: rotor type
Not specified
Pelleting: speed (g)
200000
Density gradient
Only used for validation of main results
Density medium
Type
Number of initial discontinuous layers
Lowest density fraction
Highest density fraction
Total gradient volume, incl. sample (mL)
Sample volume (mL)
Orientation
Rotor type
Speed (g)
Duration (min)
Fraction volume (mL)
Fraction processing
Pelleting: volume per fraction
Pelleting: duration (min)
Pelleting: rotor type
Pelleting: speed (g)
Pelleting-wash: volume per pellet (mL)
Pelleting-wash: duration (min)
Pelleting-wash: speed (g)
Density cushion
Density medium
Sucrose
Characterization: Protein analysis
Protein Concentration Method
microBCA
Western Blot
Detected EV-associated proteins
Alix/ NHE3/ AQP2/ CD9/ TSG101
Not detected EV-associated proteins
PKD1/ PKD2
Detected contaminants
Not detected contaminants
Albumin/ Uromodulin
Proteomics database
No
Characterization: Particle analysis
EM
EM-type
EV concentration
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