冻结破碎法提取组织蛋白方法初探

Tissue Protein Extraction by Freezing and Breaking Method

目的比较冻结破碎法和匀浆法提取组织蛋白的效率,拟建立一种提取组织蛋白的新方法。方法分别用冻结破碎器具SK-100和匀浆机处理小鼠不同部位的组织,提取总蛋白后进行蛋白定量,然后用Western blot检测β-actin蛋白表达。结果冻结破碎法虽比匀浆法耗时稍长,但与匀浆法提取的总蛋白浓度以及β-actin蛋白表达无明显统计学差异。结论匀浆法与冻结破碎法均可以用于提取小鼠各种组织的蛋白,都可以在实验室中展开,进而在蛋白质水平上进行相关功能的研究。

Objective To compare freezing and breaking,and homogenizing methods and establish a new method for total protein extraction. Methods Total protein was extracted by freezing and breaking apparatu（sSK-100）and homogenizer respectively.Then,the protein concentration was measured and subjected to Western-blot to examine β-actin expression. Results Although the consuming time of freezing and breaking approach was comparatively longer than that of homogenizing method,there was no difference in concentration of total protein and β-actin expression between both extraction methods. Conclusion Total protein can be extracted by both freezing and breaking approach and homogenizing method. The two kinds of methods can be used in the laboratory and available for functional analysis of proteins.