Preparation,identification and tissue localization of PIWIL proteins
- VernacularTitle:PIWIL系列蛋白多克隆抗体制备、鉴定及组织分布
- Author:
Yuxia LU
;
Zhigang HUANG
;
Ximei CHEN
;
Hengjun GAO
;
Ying HU
;
Xiaoyan ZHANG
;
Xun MENG
- Publication Type:Journal Article
- Keywords:
argonaute protein;
polyclonal antibody;
tissue chip
- From:
Basic & Clinical Medicine
2006;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare polyclonal antibodies against human PIWIL and to identify their property and tissue distribution of PIWIL.Methods PIWIL polypeptide was synthesized and conjugated to Keyhole limpet hemocyanin(KLH) as an immunogen.Then PIWIL-KLH conjugations were injected into rabbits subcutaneously to produce polyclonal antibodies.The specificity and sensitivity of antibodies were identified by ELISA and Western blot after purification by affinity chromatography.PIWIL were then stained on the tissue chip to study their distribution.Results Rabbit antibodies against PIWIL were prepared after injection of PIWIL-KLH conjugation.These antibodies specially recognized PIWIL peptides.Expression of PIWIL was found in the cytoplasm of epithelia cells of varied normal tissues and tumor tissues.Conclusion The successful preparation of the polyclonal antibody against PIWIL will provide an efficient reagent for further study of its role in the pathway of miRNA and RNA interference and in the pathogenesis of human disease.
