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Optimization of cell lysis and protein digestion protocols for the analysis of HeLa S3 cells by LC‐MS/MS
Author(s) -
Winter Dominic,
Steen Hanno
Publication year - 2011
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201100162
Subject(s) - lysis , hela , digestion (alchemy) , cell , chemistry , chromatography , lysis buffer , microbiology and biotechnology , biology , biochemistry
Abstract In order to maximize the number of proteins identified from Hela S3 cell lysate we tested various cell lysis, protein precipitation and digestion protocols. First, we compared three different lysis buffers, two mechanical cell disruption methods and two precipitation methods. Then, we tested six different in‐solution digestion protocols, three different in‐gel digestion protocols and ten different peptide extraction protocols. The result is a proposal for an optimized protocol to prepare the whole cell lysate samples from HeLa S3 cells.