摘要
目的寻找提取小鼠组织和细胞中组蛋白的简易方法。方法用RIPA裂解法(A法)、酸提法(B法)和组蛋白提取试剂盒法(C法)提取小鼠肝脏和A549细胞中组蛋白,用考马斯亮蓝染色及免疫印迹法检测组蛋白和乙酰化组蛋白。结果考马斯亮蓝染色结果发现B法和C法条带较A法清晰,且免疫印迹结果表明B法和C法中H3和H4条带面积更大。结论酸提法和组蛋白提取试剂盒法提取的组蛋白种类和纯度都明显高于RIPA裂解法,但酸提法成本较低、简单易行。
Objective To look for a simple method for extracting the histones from mouse tissues and cells. Methods The histones were extracted from mouse liver and A549 cells by RIPA lysis（A）, acid extraction（B） and Histone extraction kit（C）. Levels of histones and acetylated histones were determined by Coomassie brilliant blue（CBB） stain and Western blot. ResultsCompared with A method, CBB stain showed the protein bands were more distinguishable, and Western blot revealed the band area of H3 and H4 was larger in B and C methods. Conclusion Acid extraction and Histone extraction kit are superior to RIPA lysis for histone extraction, but the former is more simple and inexpensive.
出处
《广东医学院学报》
2015年第5期538-541,共4页
Journal of Guangdong Medical College
基金
国家自然科学基金(No.81170327)
广东省自然科学基金(No.9252402301000002)
东莞市医疗卫生单位科技计划一般项目(No.2012108 102022)
广东医学院建博科技创新团队项目(No.STIF201102)
关键词
组蛋白
乙酰化组蛋白
酸提法
histone
acetylated histone
acid extraction
