VISUALIZATION OF PROTEINS WITH A SILVER STAIN - A CRITICAL ANALYSIS
VISUALIZATION OF PROTEINS WITH A SILVER STAIN - A CRITICAL ANALYSIS
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DOI：
10.1002/elps.1150020304
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发表时间：
1981-01-01
期刊：
影响因子：
2.9
通讯作者：
NEUHOFF, V
中科院分区：
文献类型：
--
作者：
POEHLING, HM;NEUHOFF, V
聚丙烯酰胺凝胶中蛋白质的最佳银染色是蛋白质上的银沉积与背景染色水平之间的精细平衡。可再现的染色需要对程序中的许多步骤进行经验标准化。与考马斯蓝不同，本形式的银染色不化学计量地染色蛋白质。染色强度与蛋白质质量不呈线性关系。通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳分离的银和考马斯染色标记蛋白的密度评价证明了这一点。尽管银染色比考马斯亮蓝染色更敏感，但敏感性的增加对于单个蛋白质是不同的，并且取决于蛋白质浓度。尽管如此，如果只有最少量的蛋白质可用，并且只需要定性或半定量评价，则它是一维和二维电泳的极好方法。
Optimal silver staining of proteins in polyacrylamide gels is a fine balance between silver deposition on the proteins and the level of background staining. Reproducible staining requires empirical standardization of many steps in the procedure. Silver staining in the present form does not stoichiometrically stain proteins, unlike Coomassie Blue. Staining intensity does not bear a linear relationship to protein mass. This is demonstrated by densitometric evaluation of both silver and Coomassie-stained marker proteins fractionated by sodium dodecyl sulfate polyarcylamide gel electrophoresis. Although silver staining is more sensitive than Coomassie Blue, the increment in sensitivity is both different for individual proteins and dependent on protein concentration. Nevertheless it is an excellent method for 1 and 2-dimensional electrophoresis if only minimal amounts of protein are available and only a qualitative or semi-quantitative evaluation is required.