Aim: To prepare a rabbit anti-human apurinic/apyrimidinic endonuclease polyclonal antibody and then have its characterization analyzed.
Methods: A rabbit polyclonal antibody was prepared through a modified rapid immune procedure and then it was purified using a specific avidity column. The efficacy of this antibody was detected by ELISA, Western blot and immunohistochemistry.
Results: The titer of the antiserum determined by ELISA was up to 1:128,000. The kaff value of the antibody was 8.96 x 10(-6) mol/L. Western blot analysis showed the antibody was of high specificity. The final purified antibody was highly specific to native form of APE1 protein in mice and rats.
Conclusion: The rabbit anti-human APE1 polyclonal antibody with high titer and specificity has been successfully prepared. It can be used not only to elucidate the roles of APE1 protein in many important cellular procedures, but also to detect the expression of the APE1 protein in mice and rats.