Preparation and identification of monoclonal antibody against angiopoietin-related protein 2
- Author:
Shu MENG
1
Author Information
1. Department of Cardiology
- Publication Type:Journal Article
- From:
Academic Journal of Second Military Medical University
2006;27(7):781-783
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To prepare a monoclonal antibody against human angiopoietin-related protein 2 (ARP2). Methods: Human spleen ARP2 gene was obtained by RT-PCR. A pET32a-ARP2 plasmid was constructed and was incorporated into E. coli. The products were purified and were used to immunize 6-week-old BALB/c female mice. Hybridoma secreting anti-ARP2 monoclonal antibody was obtained by standard procedure. Mass production was carried out after specificity identification with Western blotting. Results: The fusion protein obtained by pET32a system had a relative molecular weight of about 570 000, which was in accordance with the theoretical value. The purity of the protein was more than 90% after purification. The antibody titer was 1: 104 in the hybridoma culture supernatant and 1: 107-108 in the ascites. The IgG2a type antibody had a relative molecular weight of about 570 000 by Western blot analysis. Immunohistochemistry method showed that the antibody bond with human ARP2. Conclusion: The prepared anti-human ARP2 monoclonal antibody in this study can be used for identification of ARP2 protein.
