| Non－decoloured In－gel Digestion of Coomassie Blue－stained Proteins |
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| 作者姓名： | ZHANGGuo-an BAOHui-min FANHui-zhi YANGPeng-yuan |
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| 作者单位： | DepartmentofChemistry,FudanUniversity,Shanghai200433,P.R.China |
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| 基金项目： | Supported by the National BasicResearch Priorities Program me of China( No.0 0 1CB5 10 2 0 2),National High- TechProgramm e of China( No.2 0 0 1AA2 30 31)andShanghai Science and Technology Developing Program me( No.0 1JC14 0 11) |
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| 摘 要： | 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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| 关 键 词： | 蛋白质 消化力 聚丙烯酰胺 凝胶 电泳 质谱分析 清蛋白 缩氨酸 |
| 收稿时间： | 2002-09-19 |
Non-decoloured In-gel Digestion of Coomassie Blue-stained Proteins |
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| ZHANGGuo-an BAOHui-min FANHui-zhi YANGPeng-yuan.Non-decoloured In-gel Digestion of Coomassie Blue-stained Proteins[J].Chemical Research in Chinese University,2003,19(4):399-403. |
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| Authors: | Zhang Guo-an BAO Hui-min FAN Hui-Zhi YANG Peng-yuan |
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| Institution: | Department of Chemistry, Fudan University, Shanghai 200433, P. R. China |
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| 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. |
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| Keywords: | In-gel digestion Polyacrylamide gel electrophoresis Coomassie brilliant blue Mass spectrometry |
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