Non-decoloured In-gel Digestion of Coomassie Blue-stained Proteins
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 re-moval of the dye before digestion, and is thus faster and saves a lot of labor. The resulted...
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Veröffentlicht in: | Chemical research in Chinese universities 2003-10, Vol.19 (4), p.399-403 |
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Format: | Artikel |
Sprache: | eng |
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Zusammenfassung: | 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 re-moval of the dye before digestion, and is thus faster and saves a lot of labor. The resulted digest canbe ana-lyzed 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. |
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ISSN: | 1005-9040 2210-3171 |