westernblot时该选择nc膜还是pvdf膜.pdfVIP

Western Blot 时该选择 NC 膜还是 PVDF 膜？ Western Blot 时最常用的两种膜是硝酸纤维素膜 （nitrocellulose，NC 膜）和 PVDF 膜（又称 Positively charged nylon）（Polyvinylidene fluoride，聚偏二氟乙烯膜）。 这两种膜各自有什么特点？我们的实验中该选择哪一种呢？来看 下面的文字。（摘自《Making and using Antibodies》） A study of the performance of nitrocellulose, mixed ester, nylon, and covalent-binding PVDF memberanes after passive protein adsorption and also after electrotransfer was done with several different proteins labeled with 125Iodine. The membranes exhibited different binding capacities in passive adsorption tests with labeled bovine serum albumin. The PVDF showed the least, and the regenerated cellulose and nylon membranes showed the most protein binding. Nitrocellulose and mixed ester membranes were midway between. In tests measuring protein retention, PVDF retained the most bound protein when washed with detergents or 5% skimmed milk. All the membranes showed virtually the same binding capacity as measured by autoradiography when tested under electrotransfer conditions with Towbins buffer. In passive adsorption tests, the membranes wxhibited a broad range of capacities but gave similar results in electrotransfer tests. These differences were ascribed to active migration of protein into the membrane matrix instead of simple diffusion and the increased hydrophobicity of Towbins transfer buffer because of the inclusion of methanol. 上面这段文字指出在被动扩散转移蛋白时，几种膜之间结合蛋白 的能力差别明显；但是当使用转膜仪转移蛋白时，各种膜之间的差别 就很小了。 The choice of membrane used for Western Blot is more critical if the blotted protein must maintain its native conformation for detection by the antibody. For example, a comparison using a guanosine triphosphate(GTP)-overlay assay showed that the activity of a bovine GTP-binding protein was barely detectable after transfer to hydrophobic PVDF membranes but was clearly detected after transfer to nitrocellulose. Western blot analysis showed the GTP-binding protein to b