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Quantitative Proteomic Analysis of Human Aqueous Humor Using iTRAQ 4plex Labeling.

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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… Click to show full 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.

Keywords: 4plex labeling; analysis; aqueous humor; itraq 4plex; human aqueous; proteomic analysis

Journal Title: Methods in molecular biology
Year Published: 2018

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