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Glaucoma pp 89–95Cite as

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

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1695))

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.

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Author information

Authors and Affiliations

  1. Bascom Palmer Eye Institute, McKnight Vision Research Center, University of Miami, 1638 N.W. 10th Avenue, Miami, FL, 33136, USA

    Anna Trzeciecka, Maria Carmen Piqueras & Sanjoy K. Bhattacharya

  2. Department of Ophthalmology, Case Western Reserve University, Cleveland, OH, 44195, USA

    Padmanabhan Pattabiraman & Carol Toris

Authors
  1. Anna Trzeciecka
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  2. Padmanabhan Pattabiraman
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  3. Maria Carmen Piqueras
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  4. Carol Toris
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  5. Sanjoy K. Bhattacharya
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Corresponding author

Correspondence to Sanjoy K. Bhattacharya .

Editor information

Editors and Affiliations

  1. Department of Ophthalmology, Massachusetts Eye and Ear Infirmary/Schepens Eye, Research Institute, Harvard Medical School, Boston, Massachusetts, USA

    Tatjana C. Jakobs

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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: Jakobs, T. (eds) Glaucoma. Methods in Molecular Biology, vol 1695. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7407-8_9

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  • DOI: https://doi.org/10.1007/978-1-4939-7407-8_9

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7406-1

  • Online ISBN: 978-1-4939-7407-8

  • eBook Packages: Springer Protocols

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