PEPTIDE AND PROTEIN MOLECULAR-WEIGHT DETERMINATION BY ELECTROPHORESIS USING A HIGH-MOLARITY TRIS BUFFER SYSTEM WITHOUT UREA
PEPTIDE AND PROTEIN MOLECULAR-WEIGHT DETERMINATION BY ELECTROPHORESIS USING A HIGH-MOLARITY TRIS BUFFER SYSTEM WITHOUT UREA
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DOI：
10.1016/0003-2697(86)90228-9
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发表时间：
1986-05-15
影响因子：
2.9
通讯作者：
GREGERSON, DS
中科院分区：
文献类型：
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作者：
FLING, SP;GREGERSON, DS
使用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳检查了各种缓冲系统解析和提供宽尺寸范围内的蛋白质和肽的分子量测定的能力。使用改良的 Laemmli 不连续缓冲液系统，在分离凝胶 (0.75 M) 和运行缓冲液 (0.05 M) 中含有高浓度 Tris，可在 1300 至 100,000 分子量范围内实现清晰的条带和高分辨率。使用或不使用不同浓度的尿素和/或甘油和/或蔗糖对线性梯度凝胶（8%至25%丙烯酰胺）进行测试。在 Tris 的高摩尔浓度下，成功进行肽电泳不需要添加尿素、甘油或蔗糖。当分别使用 0.75 和 0.05 M 的 Tris 运行时，不使用这些试剂运行的凝胶在整个分子量范围内显示出优异的分辨率，从而无需使用其他系统中使用的尿素或其他添加剂。因此，单一凝胶能够解析从大蛋白质到小肽的整个范围。
Various buffer systems were examined for their ability to resolve and provide molecular weight determinations of proteins and peptides over a wide size range using electrophoresis in sodium dodecyl sulfate-polyacrylamide gels. Sharp bands and high resolution were achieved in the 1300 to 100,000 molecular weight range using a modified Laemmli discontinuous buffer system with high concentrations of Tris in the resolving gel (0.75 M) and in the running buffer (0.05 M). Linear gradient gels (8 to 25% acrylamide) were tested with and without varying concentrations of urea and/or glycerol and/or sucrose. At this high molarity of Tris, the inclusion of urea, glycerol, or sucrose proved unnecessary for successful peptide electrophoresis. Gels run without these reagents showed superior resolution throughout the entire molecular weight range when run with Tris at 0.75 and 0.05 M, respectively, obviating the need for urea or other additives as used in other systems. A single gel is thus able to resolve an entire range from large proteins to small peptides.