Acta Anatomica Sinica ›› 2024, Vol. 55 ›› Issue (2): 237-240.doi: 10.16098/j.issn.0529-1356.2024.02.016

• Technology and Methodology • Previous Articles     Next Articles

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

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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