The silver staining procedure of sodium dodecyl sulfate‐gels may be accelerated by shortening fixation time
The silver staining procedure of sodium dodecyl sulfate‐gels may be accelerated by shortening fixation time
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DOI：
10.1002/elps.1150140109
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发表时间：
1993
期刊：
影响因子：
2.9
通讯作者：
S. Kirkeby;D. Moe;T. Bøg‐Hansen
中科院分区：
文献类型：
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作者：
S. Kirkeby;D. Moe;T. Bøg‐Hansen
在大多数银染色方法中，对十二烷基硫酸钠-聚丙烯酰胺凝胶电泳后分离的蛋白质进行染色的第一步是相当长时间的凝胶固定。最佳固定时间应较短，不引起背景染色，并有效地将蛋白质固定在凝胶中，而不会掩盖蛋白质与染色溶液的反应。此外，由于固定剂的潜在毒性，固定化合物的浓度应尽可能低。如果凝胶在乙醇-乙酸中预固定10 min，则用极低浓度的甲醛和戊二醛乙醇混合物或甲醛或戊二醛苦味酸和乙醇溶液固定仅5 min即可满足这些要求。根据这些结果，提出了快速灵敏的银染色程序。
In most silver staining methods the first step in the staining of proteins separated after sodium dodecyl sulfate‐polyacrylamide gel electrophoresis is a rather protracted fixation of the gels. Optimum fixation should be short, cause no background staining and effectively immobilize the proteins in the gel without masking the proteins for reaction with the staining solution. Further, the concentration of the fixing compounds should be as low as possible due to the potentional toxicity of fixatives. Fixation for only 5 min with mixtures of very low concentrations of formaldehyde and glutaraldehyde in ethanol, or a solution of formaldehyde or glutaraldehyde in picric acid and ethanol, fulfill these demands, provided that the gels were prefixed in ethanol‐acetic acid for 10 min. As a consequence of these results a fast and sensitive silver staining procedure is proposed.