A SENSITIVE SILVER STAIN FOR DETECTING LIPOPOLYSACCHARIDES IN POLYACRYLAMIDE GELS
A SENSITIVE SILVER STAIN FOR DETECTING LIPOPOLYSACCHARIDES IN POLYACRYLAMIDE GELS
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DOI：
10.1016/0003-2697(82)90673-x
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发表时间：
1982-01-01
影响因子：
2.9
通讯作者：
FRASCH, CE
中科院分区：
文献类型：
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作者：
TSAI, CM;FRASCH, CE
通过改进用于蛋白质的Ag染色方法，开发了一种用于检测聚丙烯酰胺凝胶中细菌脂多糖的敏感Ag染色剂。在十二烷基硫酸钠存在下，用聚丙烯酰胺凝胶电泳分析脂多糖，然后用修饰银染色或周期性酸希夫染色显示。Ag染色法将脂多糖染色成深褐色，其灵敏度是周期酸-希夫染色法的500倍，可检测到小于5 ng的粗型脂多糖。5 .亩分析。从鼠伤寒沙门氏菌和大肠杆菌O111:B4中提取的光滑型脂多糖分别含有30-40种不同分子量的组分。建议使用具有已知结构和O侧链重复单元数目可变的脂多糖，如上述2种脂多糖中的1种，作为分子量标记，用于估计其他脂多糖或其组分的分子量。脂多糖也可以被染成灰绿色，但使用基于ag的颜色染色方法，当样品负载较重时，脂多糖会变成灰蓝色。
A sensitive Ag stain for detecting bacterial lipopolysaccharides in polyacrylamide gels was developed by modifying the Ag-staining method used for proteins. Lipopolysaccharides were analyzed by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate followed by visualization with the modified Ag stain or periodic acid-Schiff stain. The lipopolysaccharides were stained dark brown by the Ag stain, which is 500 times more sensitive than the periodic acid-Schiff stain and can detect less than 5 ng of rough-type lipopolysaccharides. Analyses of 5 .mu.g of smooth-type lipopolysaccharides from Salmonella typhimurium and Eschericahia coli O111:B4 showed each to have 30-40 components of different MW. The use of a lipopolysaccharide having a known structure and variable numbers of repeating units in the O side chain, such as 1 of the 2 lipopolysaccharides mentioned above, as MW markers is proposed for the estimation of the MW of other lipopolysaccharides or their components. The lipopolysaccharides can also be stained grayish green, but become grayish blue with a heavy sample load, using a Ag-based color-staining method.