Antibodies specific for D-myo-inositol 1, 4, 5-trisphosphate and the enzyme-linked immunosorbent assay of D-myo-inositol 1, 4, 5- trisphosphate

摘要

Antibodies against Ins(1,4,5)P.sub.3 were raised by immunizing rabbits with two types of IP.sub.3 -BSA conjugates which were synthesized by covalently coupling Ins(1,4,5)P.sub.3 to the carrier protein via alkyl linkages. The anti-Ins(1,4,5)P.sub.3 antibodies were detected by an ELISA where Ins(1,4,5)P.sub.3 was covalently attached to a microplate well surface. Both antiserum preparations showed specific binding with Ins(1,4, 5)P.sub.3. The specificity of these antibodies was enhanced by affinity purification for the antiserum through Ins(1,4,5)P. sub.3 -agarose chromatography. The affinity-purified antibodies have IC. sub.50 values of 12 nM and 73OnM for Ins(1,4,5)P.sub.3 and Ins(1,3,4,5)P. sub.4, respectively. These antibodies showed many properties similar to those of biologically relevant receptors for Ins(1,4,5)P.sub.3.