蛋白质印迹法中免染技术和总蛋白定量方法的研究进展  

Progress of Stain-Free Technology and Total Protein Analysis in Western Blotting

作　　者：韩欣霖 杨佳儒 宝福凯 柳爱华[1,4,5,3,6] HAN Xin-Lin;YANG Jia-Ru;BAO Fu-Kai;LIU Ai-Hua(Yunnan Province Key Laboratory for Tropical Infectious Diseases in Universities,Kunming Medical University, Kunming 650500,China;Institute for Tropical Medicine,Kunming Medical University, Kunming 650500,China;Department of Microbiology and Immunology,Kunming Medical University, Kunming 650500,China;Department of Biochemistry and Molecular Biology,Kunming Medical University, Kunming 650500,China;Yunnan Province Integrative Innovation Center for Public Health, Diseases Prevention and Control, Kunming 650500,China;Yunnan Demonstration Base of International Science and Technology Cooperation for Tropical Diseases, Kunming 650500,China)

机构地区：[1]昆明医科大学云南省高校热带传染病重点实验室,云南昆明650500 [2]昆明医科大学病原生物学与免疫学系,云南昆明650500 [3]昆明医科大学生物化学与分子生物学系,云南昆明650500 [4]云南省公共卫生与疾病防控协同创新中心,云南昆明650500 [5]昆明医科大学热带医学研究所,云南昆明650500 [6]云南省热带病示范型国际科技合作基地,云南昆明650500

出　　处：《生物技术通讯》2017年第5期709-714,共6页Letters in Biotechnology

基　　金：国家自然科学基金(81371835;31560051;81560596)

摘　　要：当含有蛋白质的SDS-PAGE胶浸泡在三氯乙酸或氯仿中时,经三氯化合物的作用,使得蛋白质中的色氨酸经紫外光照射会发出可见光,因而可在SDS-PAGE凝胶上实现对蛋白质条带的定位与定性。免染技术即是基于此原理而发明的。基于免染技术,我们可以检测到待测样品中的总蛋白条带的光密度,进而利用目的蛋白条带光密度与总蛋白条带光密度的比值对目的蛋白进行定量。本文就蛋白质印迹法技术中的免染技术和总蛋白定量方法的原理和优势进行简要综述。Based on the principle that after UV stimulation,the tryptophan will emit visible light when a SDSPAGEgel containing proteins be soaked in the trichloroacetic acid or chloroform,the stain-free technology was invented.By means of stain-free technology,we can examine the optical density(OD)value of total protein in samples and then,we can quantify the target protein through utilizing the OD value ratios between target protein andtotal proteins.Here,we summarized the principle and advantages of the stain-free technology and the total proteinanalysis method.

关 键 词：免染技术 β肌动蛋白(β-actin) β微管蛋白(β-tublin) 甘油醛3-磷酸脱氢酶(GAPDH) 总蛋白定量 

分 类 号：Q51[生物学—生物化学]