Dorri Yaser, Kurien Biji T, Scofield R Hal
Arthritis and Immunology Program, Oklahoma Medical Research Foundation, Oklahoma, USA.
J Biomol Tech. 2010 Apr;21(1):1-2.
Two-dimensional gel electrophoresis (2DE) and SDS-PAGE are the two most useful methods in protein separation. Proteins separated by 2DE or SDS-PAGE are usually transferred to membranes using a variety of methods, such as electrophoretic transfer, heat-mediated transfer, or nonelectrophoretic transfer, for specific protein detection and/or analysis. In a recent study, Pettegrew et al. claim to reuse transfer buffer containing methanol for at least five times for transferring proteins from SDS-PAGE to polyvinylidene difluoride. They add 150-200 ml fresh transfer solution each time for extended use as a result of loss of transfer buffer. Finally, they test efficiency of each protein transfer by chemiluminescence detection. Here, we comment on this report, as we believe this method is not accurate and useful for protein analysis, and it can cause background binding as well as inaccurate protein analysis.
二维凝胶电泳（2DE）和十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）是蛋白质分离中最有用的两种方法。通过2DE或SDS-PAGE分离的蛋白质通常使用多种方法转移到膜上，如电泳转移、热介导转移或非电泳转移，用于特定蛋白质的检测和/或分析。在最近的一项研究中，佩特格鲁等人声称将含有甲醇的转移缓冲液重复使用至少五次，用于将蛋白质从SDS-PAGE转移到聚偏二氟乙烯膜上。由于转移缓冲液的损失，他们每次添加150 - 200毫升新鲜转移溶液以延长使用时间。最后，他们通过化学发光检测来测试每次蛋白质转移的效率。在此，我们对该报告发表评论，因为我们认为这种方法对于蛋白质分析不准确且无用，并且它会导致背景结合以及蛋白质分析不准确。
