title: Usage of electrostatic eliminator reduces human keratin contamination significantly in gel-based proteomics analysis.
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.
authors.affiliation: Department of Structural Pathology, Institute of Nephrology, Graduate School of Medical and Dental Sciences, Niigata University, Niigata, Japan. kyonami@med.niigata-u.ac.jp
authors.forename: Bo
authors.initials: B
authors.lastname: Xu
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
authors.forename: Zongjiang
authors.initials: Z
authors.lastname: Zhao
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Yoshida
authors.forename: Sameh
authors.initials: S
authors.lastname: Magdeldin
authors.forename: Hidehiko
authors.initials: H
authors.lastname: Fujinaka
authors.forename: Tamer Ahmed
authors.initials: TA
authors.lastname: Ismail
authors.forename: Eishin
authors.initials: E
authors.lastname: Yaoita
authors.forename: Tadashi
authors.initials: T
authors.lastname: Yamamoto
mesh_terms: D000818:Animals; D004340:Drug Contamination / Q000517:prevention & control*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D007633:Keratins / Q000032:analysis*; D011506:Proteins / Q000032:analysis; D040901:Proteomics / Q000379:methods; D051381:Rats; D013194:Staining and Labeling; D055672:Static Electricity
chemical_list: D011506:Proteins; D007633:Keratins