核因子-κB电泳迁移率变动分析方法的改进

A Modified Electrophoretic Mobility Shift Assay For Nuclear Factor-kappa B

目的:建立一种快速、简化的电泳迁移率变动分析(EMSA)方法检测DNA结合蛋白核因子-κB(NF-κB)。方法:NF-κB的提取和浓度测定,32P标记探针,探针与核蛋白结合,电泳和放射自显影。结果:该法可获得背景干净、条带清晰整齐的图片,能直接目测NF-κB的活性。结论:改良的EMSA用于检测NF-κB更简便、实用、敏感性高,值得推广。

Objective： To establish a fast and simple method of electrophoretic mobility shift assay（ EMSA）for the iden tification of DNA-binding protein nuclear factor-kappa B（NF-kB）. Methods： NF-kB protein was extracted and its concentration was determined. DNA probe was labeled by ^32 p. The probe was conjugated with NF-kB protein together and underwent electrophoresis. The X-ray film was exposed and autoradiography was obtained. Results： By using this method, highly distinct bands and clear background in the picture were obtained and the activity of NF-kB could be detected by eyes. Conclusion： The modified EMSA is more simple, feasible, and highly sensitive method for detection of DNA-binding proteins.