Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Video

Updated: Jun 12, 2026

Proteomic Profile of EPS-Urine through FASP Digestion and Data-Independent Analysis
14:48

Proteomic Profile of EPS-Urine through FASP Digestion and Data-Independent Analysis

Published on: May 8, 2021

Optimizing a proteomics platform for urine biomarker discovery.

Maryam Afkarian1, Manoj Bhasin, Simon T Dillon

  • 1Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts 02114, USA.

Molecular & Cellular Proteomics : MCP
|June 1, 2010
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Targeted removal of soluble Fms-like tyrosine kinase 1 in very preterm preeclampsia: a pilot trial.

Nature medicine·2026
Same author

Microsurgical Isolation and Molecular Characterization of the Mouse Left Internal Mammary Artery: Insights into Natural Resistance to Atherosclerosis.

bioRxiv : the preprint server for biology·2026
Same author

A single-cell atlas characterizes dysregulation of the bone marrow immune microenvironment associated with outcomes in multiple myeloma.

Nature cancer·2026
Same author

Butyrophilin 2A2 promotes T cell immunoregulation via CD45 phosphatase activation and protects against murine autoimmune glomerulonephritis and pregnancy loss.

Nature communications·2026
Same author

Pointing Care for Preeclampsia in the Right Direction.

Hypertension (Dallas, Tex. : 1979)·2025
Same author

First-in-child phase I trial of p-STAT3 inhibitor WP1066 in pediatric brain tumor patients.

JCI insight·2025
Same journal

A temporal phospho-acetylome atlas of human myogenesis identifies coordinated post-translational regulation.

Molecular & cellular proteomics : MCP·2026
Same journal

Temporal proteomic characterization of SARS-CoV-2 infected mouse lungs.

Molecular & cellular proteomics : MCP·2026
Same journal

Platelet proteome links metabolism to reactivity in Essential Thrombocythemia.

Molecular & cellular proteomics : MCP·2026
Same journal

Genetic rescue of disrupted synaptic protein interaction network dynamics following SYNGAP1 reactivation.

Molecular & cellular proteomics : MCP·2026
Same journal

ASAP-ID: Proximity labelling with small tags.

Molecular & cellular proteomics : MCP·2026
Same journal

Proteome profiling reveals NQO2 activity contributing to proteasome inhibitor resistance in multiple myeloma cell lines.

Molecular & cellular proteomics : MCP·2026
See all related articles

This study optimized urine proteomics for biomarker discovery, improving reproducibility with methanol precipitation and specific iTRAQ (isobaric tags for relative and absolute quantitation) conditions. The method shows promise for distinguishing patient groups.

Area of Science:

  • Proteomics
  • Biomarker Discovery
  • Urine Analysis

Background:

  • Urine biomarker discovery faces challenges due to poor reproducibility and lack of standardized proteomics protocols.
  • Developing reliable methods is crucial for identifying urinary biomarkers for various diseases.

Purpose of the Study:

  • To optimize a quantitative proteomics strategy for reproducible urine biomarker discovery.
  • To establish standardized protocols applicable to both fresh and frozen urine samples.

Main Methods:

  • Standardization of isobaric tags for relative and absolute quantitation (iTRAQ) using healthy urine.
  • Evaluation of factors including protease inhibitors, protein quantity, iTRAQ label amounts, extraction methods, and albumin/IgG depletion.
  • Application of the optimized protocol to frozen urine samples from diabetic patients with and without nephropathy.

More Related Videos

CRISPR-Cas-mediated Multianalyte Synthetic Urine Biomarker Test for Portable Diagnostics
04:33

CRISPR-Cas-mediated Multianalyte Synthetic Urine Biomarker Test for Portable Diagnostics

Published on: December 8, 2023

Related Experiment Videos

Last Updated: Jun 12, 2026

Proteomic Profile of EPS-Urine through FASP Digestion and Data-Independent Analysis
14:48

Proteomic Profile of EPS-Urine through FASP Digestion and Data-Independent Analysis

Published on: May 8, 2021

CRISPR-Cas-mediated Multianalyte Synthetic Urine Biomarker Test for Portable Diagnostics
04:33

CRISPR-Cas-mediated Multianalyte Synthetic Urine Biomarker Test for Portable Diagnostics

Published on: December 8, 2023

Main Results:

  • Methanol precipitation yielded the highest protein recovery and spectral reproducibility.
  • Minimum protein input of 20 μg/sample and a quarter unit of iTRAQ label were found to be optimal.
  • Albumin and IgG depletion did not enhance proteome coverage.
  • The optimized protocol showed potential in segregating diabetic patient groups based on proteomic profiles.

Conclusions:

  • An optimized quantitative proteomics protocol enhances reproducibility in urine biomarker discovery.
  • The developed method is suitable for analyzing both fresh and cryopreserved urine.
  • Further validation is needed, but the approach shows promise for identifying disease-specific urinary biomarkers.