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XB-ART-45846
Methods Mol Biol January 1, 2012; 917 391-407.

Complementary proteomic analysis of protein complexes.

Greco TM , Miteva Y , Conlon FL , Cristea IM .


Abstract
Proteomic characterization of protein complexes leverages the versatile platform of liquid chromatography-tandem mass spectrometry to elucidate molecular and cellular signaling processes underlying the dynamic regulation of macromolecular assemblies. Here, we describe a complementary proteomic approach optimized for immunoisolated protein complexes. As the relative complexity, abundance, and physiochemical properties of proteins can vary significantly between samples, we have provided (1) complementary sample preparation workflows, (2) detailed steps for HPLC and mass spectrometric method development, and (3) a bioinformatic workflow that provides confident peptide/protein identification paired with unbiased functional gene ontology analysis. This protocol can also be extended for characterization of larger complexity samples from whole cell or tissue Xenopus proteomes.

PubMed ID: 22956100
PMC ID: PMC3616387
Article link: Methods Mol Biol
Grant support: [+]

References [+] :
Fang, Affinity separation and enrichment methods in proteomic analysis. 2008, Pubmed


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