Tricine电泳优化方法在针灸效应小分子蛋白检测中的应用

Application of Optimized Tricine Electrophoresis Method in Detecting Low Molecular Weight Proteins of Acupuncture Effect

目的改进一种适合分离、鉴定针灸效应小分子蛋白的Tricine-SDS-PAGE电泳体系。方法研究调整分离胶中聚丙烯酰胺浓度和凝胶交联度并加入适量甘油,从而优化Tricine电泳系统。结果与常规Glycine-SDS-PAGE电泳系统比较,分离胶聚丙烯酰胺浓度为l6.8%、交联度为5.8%、含10.4%甘油的Tricine-SDS-PAGE电泳明显减少10 kD分子量蛋白的弥散,成功分离了分子量为11.0 kD的S100A8和11.8 kD的S100A11等两个针刺抗哮喘小分子蛋白。结论优化的Tricine-SDS-PAGE电泳系统可有效分离针灸效应小分子蛋白。

Objective To improve a Tricine-SDS-PAGE electrophoresis technology suitable for isolation and identification of low molecular weight proteins of acupuncture effect. Methods Tricine electrophoresis system was optimized by adjusting polyacrylamide concentration and the crosslinking degree in separation gel and adding an appropriate amount of glycerol. Results Compared with conventional Giycine-SDS-PAGE electrophoresis system, Tricine-SDS-PAGE electrophoresis system containing separation gel with 16.8% polyacrylamide and 5.8% crosslinking degree, and 10.4% glycerol significantly reduced 10 kD protein dispersion and successfully isolated two low molecular weight anti-asthma proteins of acupuncture effect： S 100A8 with molecular weight of 11.0 kD and S100All with molecular weight of 11.8 kD. Conclusion The optimized Tricine-SDS-PAGE electrophoresis system can effectively isolate low molecular weight proteins of acupuncture effect.