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使用表面导电玻璃作为阳极将蛋白质从聚丙烯酰胺凝胶快速电转移至硝酸纤维素膜。

Rapid electrotransfer of proteins from polyacrylamide gel to nitrocellulose membrane using surface-conductive glass as anode.

作者信息

Svoboda M, Meuris S, Robyn C, Christophe J

出版信息

Anal Biochem. 1985 Nov 15;151(1):16-23. doi: 10.1016/0003-2697(85)90046-6.

Abstract

A new type of inexpensive horizontal apparatus for the electrophoretic transfer of proteins from a gel to an immobilization membrane has been developed. In this system, gel and membrane were directly pressed between two flat electrodes. A surface-conductive glass was used as anode and a stainless-steel plate as cathode. Proteins could be transferred from polyacrylamide gel to nitrocellulose sheet, with a yield of at least 90% in 60-90 min, without overheating, using a voltage gradient of 30-40 V/cm. Moderate volumes of separate anodic (with 20% methanol) and cathodic (with 0.1% sodium dodecyl sulfate) buffers were suited for optimal transfer of proteins with relative molecular mass (Mr) in the 14,000-94,000 range.

摘要

已开发出一种新型的廉价水平装置,用于将蛋白质从凝胶电泳转移至固定膜。在该系统中,凝胶和膜直接压在两个扁平电极之间。使用表面导电玻璃作为阳极,不锈钢板作为阴极。蛋白质可以从聚丙烯酰胺凝胶转移至硝酸纤维素膜,在30 - 40 V/cm的电压梯度下,60 - 90分钟内产率至少为90%,且不会过热。适量体积的单独阳极缓冲液(含20%甲醇)和阴极缓冲液(含0.1%十二烷基硫酸钠)适合相对分子质量(Mr)在14,000 - 94,000范围内的蛋白质的最佳转移。

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