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Production of Nurr-1 Specific Polyclonal Antibodies Free of Cross-reactivity Against Its Close Homologs, Nor1 and Nur77
Published on: August 17, 2015
[Preparation and identification of monoclonal antibody against repression and activation protein 1]
Lin Meng1, Chengchao Shou1, Shenyi Lian1
1Ministry of Education Key Laboratory of Carcinogenesis and Translational Research, Laboratory of Biochemistry and Molecular Biology, Cancer Hospital & Beijing Institute for Cancer Research, Peking University, Beijing 100142, China.
Objective:
To prepare and identify monoclonal antibody (mAb) against human telomere-associated repression and activation protein 1 (Rap1).
Methods:
BALB/c mice were immunized with recombinant Rap1 protein. The spleen cells of the immunized mice were fused with Sp2/0 cells. The positive clones were confirmed and selected by indirect ELISA for titer determination and specificity identification.
Results:
One hybridoma cell strain secreting specific mAb against Rap1 was obtained, namely 6F11. ELISA showed that the titer of the ascites was 1:10 000. Western blotting, immunoprecipitation and immunofluorescence experiments demonstrated that the mAb could specifically recognize and bind Rap1.
Conclusion:
We have prepared mAb against human telomere-associated protein Rap1. The Rap1 mAb has a good binding ability and specificity.
