Analysis of rice proteins using SDS-PAGE shotgun proteomics

Karlie A. Neilson, Iniga S. George, Samantha J. Emery, Sridevi Muralidharan, Mehdi Mirzaei, Paul A. Haynes

    Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

    13 Citations (Scopus)

    Abstract

    In this chapter we describe the workflow used in our laboratory to analyze rice leaf samples using label-free shotgun proteomics based on SDS-PAGE fractionation of proteins. Rice proteomics has benefitted substantially from successful execution of shotgun proteomics techniques. We describe steps on how to proceed starting from rice protein extraction, SDS-PAGE, in-gel protein digestion with trypsin, nanoLC-MS/MS, and database searching using the GPM. Data from these experiments can be used for spectral counting, where simultaneous quantitation of several thousand proteins can be obtained.

    Original languageEnglish
    Title of host publicationPlant proteomics
    Subtitle of host publicationMethods and protocols
    EditorsJesus V. Jorrin-Novo, Setsuko Komatsu, Wolfram Weckwerth, Stefanie Wienkoop
    Place of PublicationNew York
    PublisherHumana Press Inc.
    Pages289-302
    Number of pages14
    Edition2
    ISBN (Electronic)9781627036313
    ISBN (Print)9781627036306
    DOIs
    Publication statusPublished - 2014

    Publication series

    NameMethods in Molecular Biology
    Volume1072
    ISSN (Print)1064-3745

    Keywords

    • Label-free
    • Plant proteomics
    • Rice
    • SDS-PAGE
    • Shotgun proteomics

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