解剖学报

解剖学报 ›› 2024, Vol. 55 ›› Issue (2): 237-240.doi: 10.16098/j.issn.0529-1356.2024.02.016

• 技术方法 • 上一篇    下一篇

水通道蛋白9商品化抗体和自制抗体识别位点的分析和应用

程全成1 丁慧如1 王子元1 方金玉1 张晓丽2* 张卫光1*   

  1. 1.北京大学基础医学院人体解剖学与组织学胚胎学系，北京 100191; 2.北京卫生职业学院医学基础教研室，北京 102402
  • 收稿日期:2023-02-15 修回日期:2023-03-27 出版日期:2024-04-06 发布日期:2024-04-06
  • 通讯作者: 张晓丽 张卫光 E-mail:zhangwg@bjmu.edu.cn
  • 基金资助:
    北京市自然科学基金

Analysis of recognition sites and application for commercial and homemade antibodies to aquaporin 9

CHENG  Quan-cheng1  DING  Hui-ru1  WANG  Zi-yuan1  FANG  Jin-yu1  ZHANG  Xiao-li2*  ZHANG  Wei-guang1*   

  1. 1.Department of Anatomy, Histology and Embryology, School of Basic Medical Sciences, Peking University Health Science Center, Beijing 100191, China; 2.Department of Preclinical Medicine, Beijing Health Vocational College, Beijing 102402, China
  • Received:2023-02-15 Revised:2023-03-27 Online:2024-04-06 Published:2024-04-06
  • Contact: ZHANG Xiao-li ZHANG Wei-guang E-mail:zhangwg@bjmu.edu.cn

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摘要：

目的 分析水通道蛋白9 (AQP9)商品化抗体和自制抗体的抗原识别位点，并鉴别其应用效果。  方法 利用Western blotting对比3种商品化抗体与自制AQP9抗体鉴定基因型的效果。分析4种抗体的抗原识别位点，及其在实际应用中的特异性。  结果 Western blotting结果显示，3种商品化抗体在野生型（WT）和Aqp9-/-小鼠中均可见蛋白条带。3种商品化抗体对应的血蓝蛋白（KLH）共轭合成肽依次为大鼠、人源和人源，与小鼠AQP9氨基酸序列存在一定差异。AQP3/7与AQP9分子量相近，且在肝中均有表达，同源性较高。自制抗体在WT鼠的27 kD位置可见一明显条带，但是Aqp9-/-鼠的相应位置未见条带。  结论 1号和3号商品化抗体可用于辅助鉴别Aqp9-/-鼠的基因型，自制抗体在蛋白水平上可以准确鉴定基因型。

关键词: 水通道蛋白9, 肝脏, 抗体, 基因敲除, 免疫印迹法, 小鼠, 大鼠, 人

Abstract:

Objective To analyze the antigen recognition sites of commercial and homemade antibodies against aquaporin (AQP) 9 ，and to identify the application effect.     Methods Western blotting was used to compare the efficacy of three commercial antibodies and self-made antibody in identifying AQP9 genotypes. The antigen recognition sites of four antibodies and their specificities in practical applications were analyzed.   Results Western blotting showed that protein bands of three commercial antibodies were detected in both WT and Aqp9-/- mice. The keyhole limpet hemocyanin(KLH) conjugated synthetic peptides corresponding to the three commercial antibodies were derived from rat, human and human, respectively. And The sequences of these three synthetic peptides were different from those of mice. AQP3/7 and AQP9 have similar molecular weight and were expressed in the liver with high homology. An obvious band of self-made antibody was observed at the 27 kD position in WT mice, but no band was observed at the corresponding position in Aqp9-/- mice.   Conclusion Commercial antibodies 1 and 3 can be used to assist in the identification of genotypes in Aqp9-/- mice. Homemade antibodies can accurately identify genotypes at the protein level.

Key words: Aquaporin 9, Liver, Antibody, Gene knockout, Western blotting, Mouse, Rat, Human

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