[Preparation and characterization of rabbit anti-human apurinic/apyrimidinic endonuclease(APE1) polyclonal antibody].
OBJECTIVE
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.


CONCLUSIONS
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.
Cao
Xiao-jing
XJ
Cancer Center, Institute of Surgery Research, Daping Hospital, Third Military Medical University, Chongqing 400042, China. caoxiaojing2006@126.com
Dai
Nan
N
Yi
Wei-jing
WJ
Li
Zeng-peng
ZP
Yang
Yu-xin
YX
Wang
Dong
D
Hu
Chuan-min
CM
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D015153:Blotting, Western; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase / Q000276:immunology* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D007106:Immune Sera / Q000276:immunology*; D008099:Liver / Q000201:enzymology; D008297:Male; D051379:Mice; D011817:Rabbits; D051381:Rats; D014599:Uterus / Q000201:enzymology
D000911:Antibodies, Monoclonal; D007106:Immune Sera; C085108:APEX1 protein, human; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase