Lobert S, Correia J J
School of Nursing, Department of Biochemistry, University of Mississippi Medical Center, Jackson 39216.
Electrophoresis. 1994 Jul;15(7):930-1. doi: 10.1002/elps.11501501135.
We present here a protocol for unidirectional wet electrotransfer of isoelectric focusing (IEF) gels to polyvinylidene difluoride (PVDF) that can be carried out in < 2 h, including preequilibration steps. In developing this protocol, we systematically evaluated sodium dodecyl sulfate and methanol concentrations in the preequilibration washes and length of electrotransfer. We demonstrated that the method is useful for tubulin, a protein that resists usual semi-dry transfer procedures due to its limited solubility under IEF conditions. We also successfully used our method for transferring the microtubule-associated protein, tau, as well as carbamylyte standards, creatine phosphokinase and carbonic anhydrase.
我们在此介绍一种用于将等电聚焦（IEF）凝胶单向湿法电转移至聚偏二氟乙烯（PVDF）的方法，该方法包括预平衡步骤，可在2小时内完成。在制定此方法时，我们系统地评估了预平衡洗涤液中的十二烷基硫酸钠和甲醇浓度以及电转移时间。我们证明该方法对于微管蛋白很有用，微管蛋白由于在IEF条件下溶解度有限，通常难以通过半干法转移。我们还成功地将该方法用于转移微管相关蛋白tau以及氨基甲酰化标准品、肌酸磷酸激酶和碳酸酐酶。
