[1]黄 震,迟秀文,韦庆文,等.Galectin-7多克隆抗体的制备与鉴定[J].现代检验医学杂志,2015,30(04):60-62.[doi:10.3969/j.issn.1671-7414.2015.04.016]
　HUANG Zhen,CHI Xiu-wen,WEI Qing-wen,et al.Preparation and Characterization of Polycolnal Antibody of Galectin-7[J].Journal of Modern Laboratory Medicine,2015,30(04):60-62.[doi:10.3969/j.issn.1671-7414.2015.04.016]
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Galectin-7多克隆抗体的制备与鉴定()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第30卷
期数:
2015年04期
页码:
60-62
栏目:
论著
出版日期:
2015-08-10

文章信息/Info

Title:
Preparation and Characterization of Polycolnal Antibody of Galectin-7
文章编号:
1671-7414(2015)04-060-04
作者:
黄 震1; 迟秀文2; 韦庆文1; 束振华1; 孙俊生1; 王 前3
1.深圳市龙岗中心医院,广东深圳 518116;
2.广东医学院护理学院,广东东莞 523808;
3.南方医科大学南方医院,广州 510515
Author(s):
HUANG Zhen1; CHI Xiu-wen2; WEI Qing-wen1; SHU Zhen-hua1; SUN Jun-sheng1; WANG Qian3
1.Longang Central Hospital of Shenzhen,Guangdong Shenzhen 518116,China;
2.Nursing School of Guangdong Medical College,Guangdong Dongguan 523808,China;
3.Nanfang Hospital of Nanfang Medical University,Guangzhou 510515,China
关键词:
半乳糖凝集素7;  表达;  抗体;  制备
分类号:
R392-33
DOI:
10.3969/j.issn.1671-7414.2015.04.016
文献标志码:
A
摘要:
目的 制备鼠抗重组半乳糖凝集素7(Galectin-7)多克隆抗体及其在膀胱癌组织中特异性鉴定。方法 PCR法从人包皮组织中扩增Galectin-7基因,并构建到pET28a表达载体中,使用IPTG诱导重组蛋白的表达,通过镍柱亲和层析纯化获得Galectin-7蛋白。以纯化的Galectin-7蛋白混合福氏完全佐剂免疫Balb/C小鼠制备抗体,并用ELISA,Western blot及免疫组织化学法检测抗体。结果 成功表达、纯化了Galectin-7重组蛋白,ELISA检测Galectin-7蛋白免疫的小鼠血清效价为1:32 000,Western blot显示该抗体能与天然的Galectin-7特异结合,免疫组织化学法检测结果表明该抗体识别人膀胱肿瘤组织的天然Galectin-7。结论 通过制备重组Galectin-7蛋白为免疫原,免疫家兔,成功地制备了效价高、特异性好的抗Galectin-7多克隆抗体。
Abstract:
Objective To prepare the mouse anti recombinanthuman Galectin-7 antibody and the antibody was characterized in bladder cancer.Methods The gene coding for Galectin-7 was amplified by PCR from the cDNA of human foreskin cells and cloned into prokaryotic expression vector pET28a.Then the recombinant plasmid pET28a/Galectin-7 was transformed into E.coli BL21(DE3)and expressed under IPTG induction.The recombinant Galectin-7 was purified through Ni2+-NT agarosegel column and the purifiedGalectin-7 used as imunogen to imunize the mouse.The titer and specificity ofthe anti- Galectin-7antibody from the mouse were analyzed by ELISA,Western blot and immunohistochemistry,respectively.Results The recombinant Galectin-7 was successfully expressed and purified,and the polyclonal ani-Galectin-7 antibody was successfully prepared.The titer of the antiserum was 1:32 000 by ELISA.Western blot analysis showed this antiboday reacted specifically with Galectin-7.Immunohistochemistry analysis showed the antibodycould recognize the native Galectin-7 in the human bladder cancer tissue.Conclusion The preparation recombinant Galectin-7 protein wasas immunogen in rabbits.It was successful to produce high titer and high specificity of anti Galectin-7 polycolonal antibody.

参考文献/References:

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备注/Memo

备注/Memo:
作者简介:黄震(1976-),男,在职博士,副主任检验师。主要从事临床免疫检验研究,Tel:13425164068,E-mail:huangzh1976@126.com。
通讯作者:王 前(1963-),男,博士生导师,主要从事临床检验教育研究。
更新日期/Last Update: 2015-08-10