[1]方莉)△,陈雪梅),任秀花),等.增强型绿色荧光蛋白兔多克隆抗体的制备[J].郑州大学学报(医学版),2010,(01):87-89.
 FANG Li),CHEN Xuemei),REN Xiuhua),et al.Preparation of polyclonal antibody of enhanced green fluorescent protein in rabbits[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2010,(01):87-89.
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增强型绿色荧光蛋白兔多克隆抗体的制备()
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2010年01期
页码:
87-89
栏目:
论著
出版日期:
2010-01-30

文章信息/Info

Title:
Preparation of polyclonal antibody of enhanced green fluorescent protein in rabbits
作者:
方莉1)陈雪梅2)任秀花2)闫志勇2)臧卫东2)
1)郑州大学医院检验科郑州4500522)郑州大学基础医学院人体解剖学教研室郑州450001
Author(s):
FANG Li1) CHEN Xuemei2) REN Xiuhua2) YAN Zhiyong2) ZANG Weidong2)
1)Clinical Laboratory, Zhengzhou University Hospital, Zhengzhou 4500522)Department of Human Anatomy, College of Basic Medical Sciences,Zhengzhou University, Zhengzhou 450001
关键词:
增强型绿色荧光蛋白多克隆抗体
Keywords:
enhanced green fluorescent protein polyclonal antibodyrabbit
分类号:
R392
文献标志码:
A
摘要:
制备增强型绿色荧光蛋白(EGFP)的兔多克隆抗体。方法:用谷胱甘肽增强型绿色荧光蛋白(GSTEGFP)融合抗原加福氏完全佐剂背部皮内注射首次免疫新西兰大白兔,第28天用GSTEGFP融合抗原加福氏不完全佐剂同样剂量加强免疫,第35天时再次免疫。第49天心脏采血30mL,4℃静置过夜,收集血清,用酶联免疫吸附试验(ELISA)检测抗血清中EGFP多克隆抗体的效价;免疫印迹法检测EGFP多克隆抗体的特异性;利用荧光免疫细胞化学检测该抗体和观察转染EGFP贴壁生长的大鼠脑皮层混合培养细胞中EGFP的表达。结果:ELISA检测EGFP多克隆抗体滴度最高为19549;免疫印迹检测结果显示该抗体特异性高;荧光免疫细胞化学检测显示在混合培养细胞中EGFP呈阳性表达。结论:直接使用GSTEGFP融合蛋白免疫新西兰大白兔,可在较短时间内制备大量EGFP多克隆抗体。
Abstract:
To prepare the polyclonal antibody of enhanced green fluorescent protein (EGFP) in rabbits.Methods:The New Zealand white rabbits were immunized with Freund’s complete adjuvant plus GSTEGFP fusion antigen at the back skin intradermal injection for the first time.The same dose Freund’s incomplete adjuvant plus GSTEGFP was injected to strengthen the immune on the 28th day and the 35th day separately. On the 49th day,blood sample was collected from heart and antisera was extracted after 4 ℃ settlement overnight. The titer of antisera was detected by ELISA. The specificity of EGFP polyclonal antibody was observed by Western blot. The expression of EGFP was examined by EGFP polyclonal antibody through fluorescence immunocytochemistry in the EGFP transfected rat cerebral cortex cells.Results:EGFP polyclonal antibody titer was up to 19 549 by ELISA. Western blot showed that the antibody had a high specificity. The expression of EGFP was positive in the mixed culture cells by fluorescence immunocytochemistry.Conclusion:EGFP polyclonal antibody can be produced in large quantities in a short time by using GSTEGFP fused protein to immunize the New Zealand white rabbit directly.

参考文献/References:

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备注/Memo

备注/Memo:
△女,1969年生,本科,主管技师,研究方向:神经疾病,Email:qrsy327@163.com
更新日期/Last Update: 2010-04-26