Efficacy of passive transfer of proteins from preparative polyacrylamide gels to nitrocellulose membranes.
Efficacy of passive transfer of proteins from preparative polyacrylamide gels to nitrocellulose membranes.
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蛋白质从制备型聚丙烯酰胺凝胶被动转移至硝酸纤维素膜的功效。
DOI：
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发表时间：
1995
影响因子：
1.4
通讯作者：
M. K
中科院分区：
文献类型：
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作者：
S. Khurana;M. K
研究了蛋白质从制备型聚丙烯酰胺凝胶到硝酸纤维素膜（NTC）的被动转移。在不从变性凝胶中除去十二烷基硫酸钠的情况下进行转移。转移是快速的，易于执行，并且不需要任何设备。通过连续转移至多个NTC，可以获得相同条带的多个复制品，而不会显著影响凝胶中剩余蛋白条带的考马斯亮蓝（CBB）染色能力。在没有电流的情况下，从低至1.5微克的蛋白质每条带在2毫米厚的凝胶被动转移的蛋白质的一部分被发现足以检测与酰胺黑染色，更是如此通过免疫染色。发现该方法对本研究中检查的7种蛋白质（14 kDa至66 kDa）非常满意，在检查其对任何蛋白质的有效性后，可用于常规定性检测。
Passive transfer of proteins from preparative polyacrylamide gels to nitrocellulose membrane (NTC) was investigated. The transfer was carried out without the removal of sodium dodecyl sulphate from the denaturing gels. The transfer was quick, easy to perform, and does not require any apparatus. More than one replica of the same band can be obtained by successive transfer to more than one NTC without significantly affecting the Coomassie Brilliant Blue (CBB) staining ability of protein bands left in the gel. A fraction of protein transferred passively in the absence of electrical current from as low as 1.5 micrograms of protein per band in a 2 mm thick gel was found sufficient for detection with amido black staining, and more so by immunostaining. The method was found highly satisfactory for seven proteins (14 kDa to 66 kDa) checked in the present study and can be adopted, after checking its efficacy for any protein, for routine qualitative tests.