兔骨组织总蛋白的提取和在免疫印迹法中的应用  

Comparison of Physical Methods of Protein Extraction in Rabbit Bone Tissue for Western Blot

作　　者：王簕[1] 裴国献[1] 

机构地区：[1]南方医科大学南方医院创伤骨科,中国广东广州510515

出　　处：《生命科学研究》2009年第2期137-141,共5页Life Science Research

基　　金：国家自然科学基金委员会-广东省联合基金(u0732003)

摘　　要：探讨快速有效的骨组织总蛋白提取法和利用该方法进行免疫印迹研究.采用单纯研磨法、单纯锤击法和锤击研磨法分别对兔骨组织蛋白进行提取,通过聚丙烯酰胺凝胶电泳对蛋白进行分离,利用免疫印迹法检测骨组织中beta-actin的表达,比较3种骨组织蛋白提取法.发现3种蛋白提取法均可满足免疫印迹研究要求,但是锤击研磨法提取的蛋白含量比单纯锤击法高,并能更好的保留49kDa以下的蛋白,而且该操作法比传统的单纯研磨法更为方便、快捷,还能节省液氮用量.锤击研磨法可作为提取骨组织蛋白的一种理想方法.To establish an efficient and quick physical method for extraction proteins from bone tissue for Western blot. Bone tissue was harvested from the femur of rabbits, and proteins were extracted by three physical methods in liquid nitrogen as follows： by simply grinding, by simply cracking with the hammer, by combination of cracking and grinding. The proteins of bone tissue were separated by SDS-PAGE, and the protein of beta-actin was assessed by Western blot. The Beta-actin of all samples can be observated by Western blot, but the concentration of proteins, expecially the molecular weight lower than 49 kDa, extracted by simple cracking with the hammer were lower than the other physical methods. Compared with simply grinding bone tissue, the method of combination of cracking and grinding was more simple and consumed less amount of liquid nitrogen.

关 键 词：骨组织 蛋白提取 免疫印迹 肌动蛋白 

分 类 号：R336[医药卫生—人体生理学]