Quantitative proteomic analysis of human aqueous humor using iTRAQ 4plex labeling

Anna Trzeciecka, Padmanabhan Pattabiraman, Maria Carmen Piqueras, Carol Toris, Sanjoy K. Bhattacharya

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Scopus citations

Abstract

iTRAQ 4plex method enables multiplexing of up to four samples for simultaneous quantitation to improve sensitivity and scope of proteomic analysis. Here, we describe iTRAQ 4plex labeling of human aqueous humor specimens followed by ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) and data analysis for peptide identification and quantitation using Proteome Discoverer software. The protocol can be applied for other animals as well; however, pooling of specimens may be required to obtain sufficient amount of protein for labeling.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages89-95
Number of pages7
DOIs
StatePublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1695
ISSN (Print)1064-3745

Keywords

  • Aqueous humor
  • Human
  • Proteins
  • Proteome
  • Relative quantitation
  • Ultra-high performance liquid chromatography-tandem mass spectrometry
  • iTRAQ 4plex

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Trzeciecka, A., Pattabiraman, P., Piqueras, M. C., Toris, C., & Bhattacharya, S. K. (2018). Quantitative proteomic analysis of human aqueous humor using iTRAQ 4plex labeling. In Methods in Molecular Biology (pp. 89-95). (Methods in Molecular Biology; Vol. 1695). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7407-8_9