[Preparation and preliminary application of rabbit anti-human PON2 antibodies(paraoxonase-2)].
OBJECTIVE
To preparation and characterize the rabbit polyclonal antibodies against human PON2 (paraoxonase-2).


METHODS
A fragment of human PON2 gene which was of low homology with rabbits but of higher hydrophilicity and immunogenicity was selected for recombinant expression in prokaryotic expression system. The rabbits were immunized with the purified GST fusion protein 3 times. The specificity and sensitivity of the anti-human PON2 polyclonal antibodies were detected by Western blot and indirect immunofluorescence.


RESULTS
The GST-PON2 fusion protein was highly expressed in Ecoli with a molecular weight of 46 kDa. Western blot analysis proved the rabbit polyclonal antibodies could specifically recognize 39 kDa native PON2 protein expressed in several cells and tissues, such as HeLa cells, U937 cells, and human liver tissue. Indirect immunofluorescence analysis confirmed that PON2 protein was located in the cytoplasm of SY5Y cells.


CONCLUSIONS
The rabbit polyclonal antibodies against human PON2 can specifically recognize natural protein expressed in human cells and tissues, Which can be used for further study and clinical detection of human PON2.
Chen
Miao
M
College of Preclinical Medicine, Anhui Medical University, Hefei 230032, China. chenmiao_6611@163.com
Yang
Jin-Ju
JJ
Li
Shu-Zhen
SZ
Liu
Xiao-Lan
XL
Liu
Ying
Y
Zhang
Lin-Jie
LJ
Gao
Jian-En
JE
Sun
Qi-Hong
QH
D000818:Animals; D000906:Antibodies / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism*; D043303:Aryldialkylphosphatase / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D015153:Blotting, Western; D002460:Cell Line; D006367:HeLa Cells; D006801:Humans; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism
D000906:Antibodies; D011993:Recombinant Fusion Proteins; D043303:Aryldialkylphosphatase; C468411:PON2 protein, human