Svoboda M, Meuris S, Robyn C, Christophe J
Anal Biochem. 1985 Nov 15;151(1):16-23. doi: 10.1016/0003-2697(85)90046-6.
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.
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。