| Effective elimination of nucleic acids from bacterial protein samples for optimized blue native polyacrylamide gel electrophoresis. | |
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MedLine Citation:
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PMID: 19598158 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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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Authors:
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Jingdan Liang; Qiule Niu; Xinping Xu; Yuanming Luo; Xiufen Zhou; Zixin Deng; Zhijun Wang |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Electrophoresis Volume: 30 ISSN: 1522-2683 ISO Abbreviation: Electrophoresis Publication Date: 2009 Jul |
Date Detail:
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Created Date: 2009-08-03 Completed Date: 2009-10-27 Revised Date: 2009-11-19 |
Medline Journal Info:
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Nlm Unique ID: 8204476 Medline TA: Electrophoresis Country: Germany |
Other Details:
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Languages: eng Pagination: 2454-9 Citation Subset: IM |
Affiliation:
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Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiaotong University, Shanghai, P. R. China. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Bacterial Proteins
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chemistry,
isolation & purification* Chemical Precipitation DNA, Bacterial / chemistry, isolation & purification, metabolism DNA-Binding Proteins / metabolism Electrophoresis, Polyacrylamide Gel / methods* Escherichia coli / chemistry Escherichia coli Proteins / chemistry Membrane Proteins / chemistry Nucleic Acids / chemistry, isolation & purification*, metabolism RNA, Bacterial / chemistry, isolation & purification, metabolism Streptomyces lividans / chemistry Streptomycin / chemistry Ultracentrifugation |
| Chemical | |
Reg. No./Substance:
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0/Bacterial Proteins; 0/DNA, Bacterial; 0/DNA-Binding Proteins; 0/Escherichia coli Proteins; 0/Membrane Proteins; 0/Nucleic Acids; 0/RNA, Bacterial; 57-92-1/Streptomycin |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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