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Effective elimination of nucleic acids from bacterial protein samples for optimized blue native polyacrylamide gel electrophoresis
Author(s) -
Liang Jingdan,
Niu Qiule,
Xu Xinping,
Luo Yuanming,
Zhou Xiufen,
Deng Zixin,
Wang Zhijun
Publication year - 2009
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200900026
Subject(s) - nucleic acid , polyacrylamide gel electrophoresis , gel electrophoresis , chemistry , chromatography , biochemistry , electrophoresis , biology , enzyme
Nucleic acids remaining within bacterial protein samples from Streptomyces lividans and Escherichia coli were found to interfere significantly with blue native polyacrylamide gel electrophoresis (BN‐PAGE), a technique used frequently for analyzing bacterial protein complexes in proteomics studies. We have used ultracentrifugation and/or precipitation of cell lysates with streptomycin sulfate to eliminate nucleic acids from total and/or membrane protein samples. Nucleic acid‐binding proteins were first enriched by precipitation with streptomycin sulfate, and contaminating nucleic acids were then eliminated by precipitation by adding polyethyleneimine. The performance of BN‐PAGE was found to be dramatically improved by these sample preparation steps.

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