Measurement of Inositol(1,4,5)Trisphosphate Using a Stereospecific Radioreceptor Mass Assay

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Abstract

Inositol(1,4,5)trisphosphate [Ins(1,4,5)P3] is an important second messenger that activates its cognate Ins(1,4,5)P3 receptor to release Ca2+ from intracellular stores. The assay described in this chapter uses the Ins(1,4,5)P3 receptor (essentially as a binding protein) to measure the biologically active trisphosphate (specifically from other trisphosphates). The binding protein (Ins(1,4,5)P3 receptor) is prepared from Bovine adrenal glands and this is mixed with [3H]-labeled and unlabeled (generated from biological samples or standards) Ins(1,4,5)P3. Using the same principles as for radioimmunoassay/ELISA the mass of Ins(1,4,5)P3 in biological samples can be estimated.