HitHunter™ Inositol (1,4,5) Triphosphate Assays
The HitHunter™ IP3 Fluorescence Polarization (FP) Assay is a rapid, homogeneous method for measuring cellular D-myo-inositol-1,4,5-trisphosphate (IP3) in a 384-well format.
Product Features
•Homogeneous FP Assay
•Robust and Sensitive Assay with EC50 < 10 nM
•Scalable assay, well-suited for HTS application
Application
•Assess Gq-coupled GPCRs
•Direct measurement of cellular inositol (1,4,5) trisphosphate (IP3)
•Detects partial agonism
Assay Principle
In the assay,cells treated with ligand to increase levels of IP3 produced are immediately quenched by perchloric acid. Fluorescent IP3 tracer and the binding protein are subsequently added. IP3 from cell lysates compete for binding to the IP3 binding protein. Bound IP3 tracer will "tumble" more slowly in solution, creating a polarized signal (high mP). The polarized signal is inversely proportional to the amount of IP3 in the cell lysates. Assay can be run on any FP plate reader. For instrument specific filters, contact the instrument manufacturer.
For instrument compatibility, please click here.
Available Collaterals
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Asay-Green
FP tracer