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

Autor:	Trzeciecka A; Bascom Palmer Eye Institute, McKnight Vision Research Center, University of Miami, 1638 N.W. 10th Avenue, Miami, FL, 33136, USA., Pattabiraman P; Department of Ophthalmology, Case Western Reserve University, Cleveland, OH, 44195, USA., Piqueras MC; Bascom Palmer Eye Institute, McKnight Vision Research Center, University of Miami, 1638 N.W. 10th Avenue, Miami, FL, 33136, USA., Toris C; Department of Ophthalmology, Case Western Reserve University, Cleveland, OH, 44195, USA., Bhattacharya SK; Bascom Palmer Eye Institute, McKnight Vision Research Center, University of Miami, 1638 N.W. 10th Avenue, Miami, FL, 33136, USA. SBhattacharya@med.miami.edu.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2018; Vol. 1695, pp. 89-95.
DOI:	10.1007/978-1-4939-7407-8_9
Abstrakt:	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.
Databáze:	MEDLINE
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