[Preparation and distribution of polyclonal antibodies against human PIWIL3 protein in tumor tissues].
OBJECTIVE
To prepare specific antibodies against human PIWIL3 protein of Argonaute family and study its expression and distribution in human tumors.


METHODS
To synthesize the polypeptide using the optimal polypeptide immunogens based on sequence homology and peptide immunogenicty in the internal polypeptide selecting database. The synthesized polypeptide were combined with KLH, then immunized the experimental rabbits to produce antiserum. The antiserum was purified by gels custodited the polypeptide using affinity chromatograph, and validated its ability to combine with the corresponding proteins by enzyme-linked immunosorbentassay (ELISA) and Western blot. The expression and distribution of PIWIL3 protein were detected by human tumor tissue biochips.


RESULTS
The specific antibodies against human PIWIL3 proteins were prepared and had high purity. PIWIL3 proteins were expressed in the cytoplasma of human astrocytic glioma and meningioma.


CONCLUSIONS
The specific antibodies against human PIWIL3 protein have been successfully prepared by using the internal polypeptide selecting database to obtain the optimal polypeptide immunogens, which are distinguished from other sequence homology proteins in the subfamily. These antibodies will have potential value for human PIWIL3 protein in human specific tumor pathogenesis.
Wang
Xiao-Lei
XL
Department of Gastroenternology, Tongji Hospital, Tongji University, Shanghai 200065, China. xlwang2006@163.com
Chen
Xi-Mei
XM
Gao
Heng-Jun
HJ
Huang
Zhi-Gang
ZG
D000906:Antibodies / Q000276:immunology*; D001254:Astrocytoma / Q000378:metabolism; D006801:Humans; D008579:Meningioma / Q000378:metabolism; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology; D011506:Proteins / Q000737:chemistry / Q000276:immunology*
D000906:Antibodies; D010455:Peptides; D011506:Proteins