Document Detail

In-gel s-pyridylethylation of gel-resolved proteins: individual spot method.
MedLine Citation:
PMID:  21356844     Owner:  NLM     Status:  In-Data-Review    
INTRODUCTIONThis protocol describes a method for performing reduction and S-alkylation of Coomassie-blue-stained proteins within individually excised, stained protein spots or bands. Reduction is performed with dithiothreitol, and alkylation with 4-vinylpyridine. (Treatment of free cysteines with 4-vinylpyridine yields the S-β-(4-pyridylethyl) cysteinyl derivative.) S-β-(4-pyridylethyl) cysteine-containing peptides can be readily identified during RP-HPLC by their characteristic absorbance at 254 nm and during electrospray ionization tandem mass spectrometry by the appearance of a characteristic pyridylethyl fragment ion of 10(6) Da. The position of cysteine residues in a polypeptide sequence can be determined either by mass spectrometry or as phenylthiohydantoin S-β-(4-pyridylethyl) cysteine during Edman degradation.
Richard J Simpson
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Publication Detail:
Type:  Journal Article     Date:  2008-06-01
Journal Detail:
Title:  CSH protocols     Volume:  2008     ISSN:  -     ISO Abbreviation:  CSH Protoc     Publication Date:  2008  
Date Detail:
Created Date:  2011-03-01     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101280522     Medline TA:  CSH Protoc     Country:  United States    
Other Details:
Languages:  eng     Pagination:  pdb.prot4605     Citation Subset:  -    
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