Mass spectrometric identification of proteins from silver‐stained polyacrylamide gel: A method for the removal of silver ions to enhance sensitivity
Mass spectrometric identification of proteins from silver‐stained polyacrylamide gel: A method for the removal of silver ions to enhance sensitivity
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DOI：
10.1002/(sici)1522-2683(19990301)20:3
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发表时间：
1999-03
期刊：
影响因子：
2.9
通讯作者：
F. Gharahdaghi;Catherine R. Weinberg;D. A. Meagher;B. Imai;S. Mische
中科院分区：
文献类型：
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作者：
F. Gharahdaghi;Catherine R. Weinberg;D. A. Meagher;B. Imai;S. Mische
质谱学是鉴定纳克级蛋白质的一种强有力的技术。然而，在质谱分析之前需要一定程度的样品准备，而银染蛋白质凝胶样品是最有问题的。在这里，我们报告了我们的策略，通过在酶消化之前脱色，从一维或二维凝胶中的银染蛋白质凝胶样品中获得多肽质量图谱。本研究表明，通过使用脱色方法，提高了基质辅助激光解吸电离飞行时间(MALDI-TOF)质谱分析的灵敏度和质量，使更多的蛋白质能够被多肽数据库分析所鉴定。
Mass spectrometry is a powerful technique for the identification of proteins at nanogram quantities. However, some degree of sample preparation prior to mass spectrometry is required, and silver‐stained protein gel samples are most problematic. Here we report our strategy to obtain peptide mass profiles from silver‐stained protein gel samples from one‐ or two‐dimensional gels by destaining prior to enzymatic digestion. This study demonstrates that by using the destaining method, the sensitivity and quality of mass spectra is increased for matrix‐assisted laser desorption ionization‐time of flight (MALDI‐TOF) mass spectrometric analysis, permitting more proteins to be identified by peptide mass database analysis.