Jägersten C, Edström A, Olsson B, Jacobson G
Pharmacia LKB Biotechnology AB, Uppsala, Sweden.
Electrophoresis. 1988 Oct;9(10):662-5. doi: 10.1002/elps.1150091007.
An improved procedure, "thermoblotting", is described for transferring proteins by diffusion from PhastGel Gradient media to an immobilizing matrix after horizontal sodium dodecyl sulfate-polyacrylamide gel electrophoresis. After electrophoresis the gels were left on the separation bed of PhastSystem, the blotting matrix was applied and a transfer temperature was selected between 5-70 degrees C. An experimental series at fixed diffusion times showed that the transfer yield was significantly increased with temperature. The evaluation was done visually after staining of the blots with colloidal gold. An evaluation study comparing nitrocellulose, nylon, and polyvinylidenedifluoride of different pore sizes is also reported. Finally, the transfer efficiencies for 125I-labelled bovine serum albumin and soybean trypsin inhibitor were estimated using four different blotting procedures: two diffusion blotting techniques and two electrophoretic blotting techniques (tank vs. semi-dry).
本文描述了一种改进的方法——“热印迹法”，即在水平十二烷基硫酸钠-聚丙烯酰胺凝胶电泳后，通过扩散将蛋白质从PhastGel梯度介质转移至固定化基质上。电泳后，将凝胶留在PhastSystem的分离床上，放置印迹基质，并在5至70摄氏度之间选择转移温度。在固定扩散时间的一系列实验表明，转移产率随温度显著增加。用胶体金对印迹进行染色后，通过目视进行评估。本文还报道了一项比较不同孔径的硝酸纤维素、尼龙和聚偏二氟乙烯的评估研究。最后，使用四种不同的印迹方法估计了125I标记的牛血清白蛋白和大豆胰蛋白酶抑制剂的转移效率：两种扩散印迹技术和两种电泳印迹技术（槽式电泳与半干电泳）。
