MISC

2008年11月

Multiplex detection and identification of proteins on a PVDF membrane blocked with a synthetic polymer-based reagent

ELECTROPHORESIS
  • Hiroshi Kawasaki
  • ,
  • Akiko Okayama
  • ,
  • Yuko Wafune
  • ,
  • Shota Yahagi
  • ,
  • Noriaki Arakawa
  • ,
  • Hisashi Hirano

29
21
開始ページ
4377
終了ページ
4380
記述言語
英語
掲載種別
DOI
10.1002/elps.200800200
出版者・発行元
WILEY-V C H VERLAG GMBH

2-DE is one of the most powerful methods for analyzing proteins expressed in cells and tissues. Immunodetection of proteins blotted on a polymer membrane is the method of choice for detecting specific proteins in 2-D gels. To precisely locate spots of immunoreactive proteins in 2-D gels, both dye staining and immunodetection were performed on the same PVDF membrane. Prior to immunodetection, nonspecific adsorption of the antibodies to the membrane was blocked with a synthetic polymer-based reagent (N-102) after protein transfer. The protein was then stained with colloidal gold or CBB followed by protein spot identification by LC-MS. Described herein is a method for multiplex analysis of proteins transferred to a PVDF membrane. Proteins that were phosphorylated at tyrosine in the phosphoproteome of rice callus or human ovarian cancer cells were detected by immunoblotting and subsequently identified with high precision.

Web of Science ® 被引用回数 : 3

リンク情報
DOI
https://doi.org/10.1002/elps.200800200
CiNii Articles
http://ci.nii.ac.jp/naid/80019992076
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/18942680
Web of Science
https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000261484100011&DestApp=WOS_CPL

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