Non-decoloured In-gel Digestion of Coomassie Blue-stained Proteins

Chemical Research in Chinese Universities ›› 2003, Vol. 19 ›› Issue (4): 399-403.

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Non-decoloured In-gel Digestion of Coomassie Blue-stained Proteins

ZHANG Guo-an, BAO Hui-min, FAN Hui-zhi, YANG Peng-yuan

Department of Chemistry, Fudan University, Shanghai 200433, P. R. China

Received:2002-09-19 Online:2003-12-24 Published:2011-08-06
Supported by:
Supported by the National Basic Research Priorities Programme of China(No.001CB510202), National High-Tech Programme of China(No.2001AA23031) and Shanghai Science and Technology Developing Programme(No.01JC14011).

Abstract

Abstract: A simplified method is presented for tryptic digestion of Coomassie brilliant blue(CBB)-stained proteins in polyacrylamide gels. Compared with conventional methods, the proposed method does not require a removal of the dye before digestion, and is thus faster and saves a lot of labor. The resulted digest can be analyzed by either RPLC/ESIMS or MALDI MS for identification of the protein in a conventional way. Model studies with bovine serum albumin(BSA) showed that 50 ng of the protein could be routinely identified. The simplified procedure displays a tendency to produce more incompletely cleaved peptides, which is favorable for improving the sequence coverage.

Key words: In-gel digestion, Polyacrylamide gel electrophoresis, Coomassie brilliant blue, Mass spectrometry

ZHANG Guo-an, BAO Hui-min, FAN Hui-zhi, YANG Peng-yuan. Non-decoloured In-gel Digestion of Coomassie Blue-stained Proteins[J]. Chemical Research in Chinese Universities, 2003, 19(4): 399-403.

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Non-decoloured In-gel Digestion of Coomassie Blue-stained Proteins

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