广西师范大学学报（自然科学版） ›› 2018, Vol. 36 ›› Issue (1): 121-125.doi: 10.16088/j.issn.1001-6600.2018.01.017

• • 上一篇    下一篇

不同固定剂对核膜蛋白免疫荧光效果的比较研究

吴卓玲1,2,3,赖艳花1,杨柳1,2,3*   

  1. 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
  • 收稿日期:2017-01-22 出版日期:2018-01-20 发布日期:2018-07-17
  • 通讯作者: 杨柳(1983—),男,河南汝南人,广西师范大学教师,博士。E-mail:yangl@gxnu.edu.cn
  • 基金资助:
    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)

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

WU Zhuoling1,2,3,LAI Yanhua1,YANG Liu1,2,3*   

  1. 1.College of Life Science,Guangxi Normal University,Guilin Guangxi 541006,China;
    2. Guangxi Universities Key Laboratory of Stem Cell and Biopharmaceutical,Guangxi Normal University,Guilin Guangxi 541004,China;3. Research Center for Biomedical Science,Guangxi Normal University,Guilin Guangxi 541004,China
  • Received:2017-01-22 Online:2018-01-20 Published:2018-07-17

PDF (PC)

681

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。

关键词: 固定剂, 免疫荧光, 核膜蛋白

Abstract: Immunofluorescence is a common method of detecting protein cell location. Different cell fixation methods may affect the staining effect. In this study, 3 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.

Key words: fixatives, immunofluorescence, nuclear membrane protein

中图分类号: 

  • R446.61
[1] BARBIERATO M,ARGENTINI C,SKAPER S D. Indirect immunofluorescence staining of cultured neural cells[J]. Methods Mol Biol,2012,836:235-246.
[2] 李冉,杨文理,覃扬,等.不同固定方法对细胞免疫荧光染色结果的影响[J].西部医学,2013,25(11):1608-1610.
[3] 范瑾瑾,骆宁,董秀清,等. 不同固定剂在激光共聚焦荧光技术中的效果评价[J].中山大学学报(医学科学版),2009,30(5):600-604.
[4] 秦犹,陈永昌.免疫荧光技术中不同固定剂对RhoA蛋白定位的影响[J]. 江苏大学学报(医学版),2006,16(5):385-388.
[5] OSTLUND C,WORMAN K J. Nuclear envelope proteins and neuromuscular diseases[J]. Muscle Nerve,2003,27(4):393-406.
[6] NAGANO A,KOGA R,OGAWA M,et al. Emerin deiciency at the nuclear membrane in patients with Emery-Dreifuss muscular dystrophy[J].Nat Genet,1996,12:254-259.
[7] SCHMIEDEBERG L,SKENE P,DEATON A,et al. A temporal threshold for formaldehyde crosslinking and fixation[J]. PLoS ONE,2009,4(2):e4636-4638.
[8] 张丽丽,耿慧霞,石贞玉,等.固定剂和传代次数对线粒体荧光质粒pDsRed2-Mito荧光强度的影响[J].河南大学学报(自然科学版),2016,46(6):670-673.
[9] 罗果,保玉心,李晋.不同固定液对细胞骨架荧光染色标记效果的影响[J].第三军医大学学报,2011,33(7):753-755.
[10] KAWAHARA A,TAIRA T,ABE H,et al.Fixation effect of SurePath preservative fluids using epidermal growth factor receptor mutation-specific antibodies for immunocytochemistry[J].Cancer Cytopathology,2014,122(2):145-152.
[11] CAETE M,JUARRANZ A,LPEZ-NIEVA P,et al. Fixation and permanent mounting of fluorescent probes after vital labelling of cultured cells[J]. Acta Histochemica,2001,103(2):117-126.
[12] WANG H,MATISE M P. Immunofluorescence staining with frozen mouse or chick embryonic tissue sections[J]. Methods Mol Biol,2013,1018:175-188.
[13] 林哲绚,罗红军,李慧,等. 免疫荧光技术中不同固定剂对细胞p65核移位观察效果的影响[J].山西医药杂志,2012,41(3): 211-213.
[14] 韩陈陈,马旸,李亦凡,等.不同固定液对内皮细胞形态和EP4荧光染色的影响[J].安徽医科大学学报,2016(9):1372-1374.
[1] 霍群, 刘杰, 陈莉, 廖维甲. 血清AFP临界值判断对肝癌诊断的影响[J]. 广西师范大学学报（自然科学版）, 2014, 32(3): 121-124.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
版权所有 © 广西师范大学学报（自然科学版）编辑部
地址：广西桂林市三里店育才路15号　邮编：541004
电话：0773-5857325　E-mail: gxsdzkb@mailbox.gxnu.edu.cn
本系统由北京玛格泰克科技发展有限公司设计开发