Nov, 2008
Multiplex detection and identification of proteins on a PVDF membrane blocked with a synthetic polymer-based reagent
ELECTROPHORESIS
- ,
- ,
- ,
- ,
- ,
- Volume
- 29
- Number
- 21
- First page
- 4377
- Last page
- 4380
- Language
- English
- Publishing type
- DOI
- 10.1002/elps.200800200
- Publisher
- 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.
- Link information
-
- 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
- ID information
-
- DOI : 10.1002/elps.200800200
- ISSN : 0173-0835
- CiNii Articles ID : 80019992076
- Pubmed ID : 18942680
- Web of Science ID : WOS:000261484100011