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Usage of electrostatic eliminator reduces human keratin contamination significantly in gel-based proteomics analysis
Faculty
Veterinary Medicine
Year:
2011
Type of Publication:
Article
Pages:
1022-1029
Authors:
Xu, Bo, Zhang, Ying, Zhao, Zongjiang, Yoshida, Yutaka, Magdeldin, Sameh, Fujinaka, Hidehiko, Ismail, Tamer Ahmed, Yaoita, Eishin, Yamamoto, Tadashi
DOI:
10.1016/j.jprot.2011.03.001
Journal:
JOURNAL OF PROTEOMICS ELSEVIER SCIENCE BV
Volume:
74
Research Area:
Biochemistry \& Molecular Biology
ISSN
ISI:000292440400013
Keywords :
Mass spectrometry, In-gel digestion, Keratin contamination, Static electricity eliminator
Abstract:
In the field of bottom-up proteomics, heavy contamination of human keratins could hinder the comprehensive protein identification, especially for the detection of low abundance proteins. In this study, we examined the keratin contamination in the four major experimental procedures in gel-based proteomic analysis including gel preparation, gel electrophoresis, gel staining, and in-gel digestion. We found that in-gel digestion procedure might be of importance corresponding to keratin contaminants compared to the other three ones. The human keratin contamination was reduced significantly by using an electrostatic eliminator during in-gel digestion, suggesting that static electricity built up on insulated experimental materials might be one of the essential causes of keratin contamination. We herein proposed a series of methods for improving experimental conditions and sample treatment in order to minimize the keratin contamination in an economical and practical way. (C) 2011 Elsevier B.V. All rights reserved.
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