Antibodies specific for d-myo-inositol 1,4,5-trisphosphate and t

Details Antibodies specific for d-myo-inositol 1,4,5-trisphosphate and t Antibodies specific for d-myo-inositol 1,4,5-trisphosphate and t

1997-10-23

2000-05-30

Scheiner, Toni R.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving antigen-antibody binding, specific binding protein...

435 71, 436543, 436547, 436528, 436532, G01N 3353

Patent

active

060689862

ABSTRACT:
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 73O nM 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.

REFERENCES:
patent: 5252707 (1993-10-01), Ozaki et al.
patent: 5393912 (1995-02-01), Chen et al.
Gon et al., Bioorg. Med. Chem, vol. 2, No. 1, pp. 7-13.
Harlow & Lane, (1988) Antibodies, A Laboratory Manual, Cold Spring Harbor Laboratory, pp. 53-88, 127-137 and 309-315.

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