Preparation and identification of monoclonal antibodies against daintain.
Daintain is a 17-kDa polypeptide originally purified from porcine intestine. This polypeptide is associated with insulin secretion and inflammatory responses. Daintain is highly similar in amino acid sequence to allograft inflammatory factor-1 (AIF-1). Here we report the preparation and identification of monoclonal antibodies (MAbs) against daintain. To enhance its immunogenicity, daintain was coupled to carrier protein bovine serum albumin (BSA) by a two-step glutaraldehyde method. Using conventional procedures, we obtained four stable hybridoma cell lines that can produce and secret anti-daintain MAbs. We further analyzed their isotypes, titer, and affinity, and found that those MAbs belong to the G1 subclass with kappa light chains. The MAbs were capable of recognizing daintain as determined by Western blotting. The produced MAbs will be a useful tool for further investigation of daintain functions in organisms.
Fu
Kang
K
Institute of Biophysics and Biochemistry, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, PR China.
Zhao
Yan-Ying
YY
Tang
Wen-Xin
WX
Zhong
Ya
Y
Yang
Tang-Bin
TB
Wang
Jian-He
JH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000302:isolation & purification; D006825:Hybridomas / Q000378:metabolism; D007074:Immunoglobulin G / Q000276:immunology*; D007422:Intestines / Q000737:chemistry; D010455:Peptides / Q000276:immunology* / Q000302:isolation & purification; D013552:Swine
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D010455:Peptides