Effective elimination of nucleic acids from bacterial protein samples for optimized blue native polyacrylamide gel electrophoresis.
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.
Liang
Jingdan
J
Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiaotong University, Shanghai, P. R. China.
Niu
Qiule
Q
Xu
Xinping
X
Luo
Yuanming
Y
Zhou
Xiufen
X
Deng
Zixin
Z
Wang
Zhijun
Z
D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification*; D011232:Chemical Precipitation; D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D004926:Escherichia coli / Q000737:chemistry; D029968:Escherichia coli Proteins / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry; D009696:Nucleic Acids / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D012329:RNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D048170:Streptomyces lividans / Q000737:chemistry; D013307:Streptomycin / Q000737:chemistry; D014461:Ultracentrifugation
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D009696:Nucleic Acids; D012329:RNA, Bacterial; D013307:Streptomycin