不同固定剂对核膜蛋白免疫荧光效果的比较研究  

Comparative Study on Effects of Fixation Nuclear Membrane Proteins by Different Fixative Agents in Immunofluorescence

作　　者：吴卓玲 赖艳花 杨柳[1,2,3] 

机构地区：[1]广西师范大学生命科学学院,广西桂林541006 [2]广西高校干细胞与医药生物技术重点实验室(广西师范大学),广西桂林541004 [3]广西师范大学生物医学研究中心,广西桂林541004

出　　处：《广西师范大学学报（自然科学版）》2018年第1期121-125,共5页Journal of Guangxi Normal University:Natural Science Edition

基　　金：广西自然科学基金(2015GXNSFCA139022);广西师范大学大学生创新创业训练计划(201510602192)

摘　　要：免疫荧光技术是检测蛋白质细胞定位的常用方法,不同的细胞固定方法可能会影响染色效果。本文以核膜蛋白lamin A/C、emerin为研究对象,利用3种不同固定剂进行免疫荧光实验。结果表明,利用体积分数为4%多聚甲醛溶液固定细胞通常可以获得良好的免疫荧光染色效果,利用甲醇固定时lamin A/C染色效果相对较弱,而利用丙酮固定的细胞其荧光信号往往呈现为核膜不连续染色。综合分析,作为最常用的固定剂,利用4%多聚甲醛溶液通常可以取得较好的染色效果;但对于新的目的蛋白来说,由于蛋白质的结构与功能特异性,适合的固定方法仍需要具体的试验检测才能确定。Immunofluorescence is a common method of detecting protein cell location, lOltterent cell fixation methods may affect the staining effect. In this study, g different fixative agents were performed for immunofluorescence experiments on nuclear membrane protein lamin A/C and emerin. It revealed good immunofluorescence staining for 4% paraformaldehyde-fixed cells, relatively weaker lamin A/C staining than emerin for methanol fixation cells, and non-continuous dyeing around nuclear membrane for the acetone-fixed cells. In an overall consideration, 4% paraformaldehyde, one of the most commonly fixative agents, can be used for good staining effect. However, the suitable fixing method for targeted new protein is still necessary to explore due to the specific protein structure and function.

关 键 词：固定剂 免疫荧光 核膜蛋白 

分 类 号：R446.61[医药卫生—诊断学] Q26[医药卫生—临床医学]