[1]郭晓姝,李洪涛,方海立,等.小鼠IFIT1多克隆抗体的制备[J].第三军医大学学报,2007,29(15):1462-1464.
 GUO Xiao-shu,LI Hong-tao,FANG Hai-li,et al.Preparation of polyclonal antibody of mouse IFIT1[J].J Third Mil Med Univ,2007,29(15):1462-1464.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
29卷
期数:
2007年第15期
页码:
1462-1464
栏目:
论著
出版日期:
2007-08-15

文章信息/Info

Title:
Preparation of polyclonal antibody of mouse IFIT1
作者:
郭晓姝李洪涛方海立王军平粟永萍
第三军医大学军事预防医学院防原医学教研室,全军复合伤研究所,创伤、烧伤与复合伤国家重点实验室;重庆医科大学基础医学院生理学教研室
Author(s):
GUO Xiao-shu LI Hong-tao FANG Hai-li WANG Jun-ping SU Yong-ping
State Key Laboratory of Trauma, Burns and Combined Injury, Institute of Combined Injury, College of Preventive Medicine, Third Military Medical University, Chongqing 400038; Department of Physiology, School of Preclinical Medicine, Chongqing Medicine University, Chongqing 400016, China
关键词:
IFIT1多克隆抗体融合蛋白抗原性免疫应答
Keywords:
IFIT1 polyclonal antibody fusion protein antigenicity immune response
分类号:
R392-33; R392.11
文献标志码:
A
摘要:
目的    获取小鼠IFIT1(Interferon-induced protein with tetratricopetide repeats 1 )特异性多克隆抗体。    方法    扩增培养高效表达IFIT1的重组子pMAL-C2X-IFIT1,超声破碎后的细胞上清过Amylose Pre-packed column亲和层析柱,将所得纯化的融合蛋白MBP-IFIT1作为抗原,添加福氏佐机剂后免疫兔,收集抗血清,用Western blot和ELISA方法测定抗血清特异性反应和效价。    结果    纯化的MBP-IFIT1可诱导兔产生特异性免疫应答,所得抗体能够特异性识别融合蛋白中的IFIT1,ELISA结果显示其效价为1∶12 800。    结论    MBP-IFIT1具有良好的抗原性,以该融合蛋白为抗原免疫兔成功制备出IFIT1特异性多克隆抗体。
Abstract:
Objective    To acquire specific polyclonal antibody of mouse IFIT1 (Interferon-induced protein with tetratricopetide repeats 1).     Methods    The single clone of pMAL-C2X-IFIT1, confirmed previously to highly express MBP-IFIT1 fusion protein, was cultivated and amplified. After ultrasonic fragmentation and centrifugation, the MBP-IFIT1 fusion protein was purified from the supernatant by using the Amylose Pre-packed column. Then, the MBP-IFIT1 fusion protein, mixed with Freund’s adjuvant, was injected into rabbits twice with two-week interval. The rabbit serum was collected, then its anti-IFIT1 titer was detected by ELISA, and its anti-IFIT1 specific reaction was detected by Western blot.     Results    The rabbit serum immunized with purified MBP-IFIT1 protein showed specific anti-IFIT1 ability, and its antibody titer was about 1∶12 800.      Conclusion    The purified MBP-IFIT1 fusion protein possesses favorable antigenicity, and highly specific polyclonal antibody of mouse IFIT1 can be obtained by using purified MBP-IFIT1 as antigen to immunize rabbit.

参考文献/References:

郭晓姝,李洪涛,方海立,等. 小鼠IFIT1多克隆抗体的制备[J]. 第三军医大学学报, 2007, 29(15):1462-1464.

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更新日期/Last Update: 2008-09-26