HitHunter™ IP3 FP Assay by DiscoveRx Corporation
Manufacturer DiscoveRx Corporation
Overview & Description Ligand binding of Gq-protein coupled receptors on the cell surface activate phopholipase C (PLC) to produce two critical second messengers – Diacylglycerol (DAG) and inositol-1,4,5-trisphosphate (IP3). IP3 moves through the cytosol and binds to the IP3 receptor in the endoplasmic reticulum to mobilize calcium stores. 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... Read more
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HitHunter™ IP3 FP Assay
Overview & Description
Ligand binding of Gq-protein coupled receptors on the cell surface activate phopholipase C (PLC) to produce two critical second messengers – Diacylglycerol (DAG) and inositol-1,4,5-trisphosphate (IP3). IP3 moves through the cytosol and binds to the IP3 receptor in the endoplasmic reticulum to mobilize calcium stores.
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. The assay is based on competitive binding between and IP3 fluorescent tracer and unlabelled IP3 from cell lysates or standards.
Features and Benefits
» Assess Gq coupled GPCRs
» Direct Measurement of cellular Inositol (1,4,5) trisphosphate (IP3)
» Faster alternative to calcium flux assays and more specific
HitHunter IP3 FP Assay Principle
The HitHunter™ IP3 FP Assay is a rapid, homogeneous method for measuring cellular D-myo-inositol-1,4,5-trisphosphate (IP3) in a 384-well format. The assay is based on competitive binding between an IP3 fluorescent tracer and unlabelled IP3 from cell lysates or standards.
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.
Product Details
Kit Contents
» IP3 Binding Protein
» IP3 tracer
» IP3 Standard
Instrument Requirements
» Fluorescence Polarization Detection
Green Tracer
Excitation: 485 nM
Emission: 530 nM
Flexible Assay Characteristics
» Homogeneous: Add and mix protocols simplify automation for quicker assays without the need for washing, centrifugation or filtering.
» Scaleable: Adaptable protocols in 96-, 384- and 1536-well applications
» Convenient: Use existing liquid handling either in batch or HTS mode with existing FP readers.
Ligand binding of Gq-protein coupled receptors on the cell surface activate phopholipase C (PLC) to produce two critical second messengers – Diacylglycerol (DAG) and inositol-1,4,5-trisphosphate (IP3). IP3 moves through the cytosol and binds to the IP3 receptor in the endoplasmic reticulum to mobilize calcium stores.
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. The assay is based on competitive binding between and IP3 fluorescent tracer and unlabelled IP3 from cell lysates or standards.
Features and Benefits
» Assess Gq coupled GPCRs
» Direct Measurement of cellular Inositol (1,4,5) trisphosphate (IP3)
» Faster alternative to calcium flux assays and more specific
HitHunter IP3 FP Assay Principle
The HitHunter™ IP3 FP Assay is a rapid, homogeneous method for measuring cellular D-myo-inositol-1,4,5-trisphosphate (IP3) in a 384-well format. The assay is based on competitive binding between an IP3 fluorescent tracer and unlabelled IP3 from cell lysates or standards.
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.
Product Details
Kit Contents
» IP3 Binding Protein
» IP3 tracer
» IP3 Standard
Instrument Requirements
» Fluorescence Polarization Detection
Green Tracer
Excitation: 485 nM
Emission: 530 nM
Flexible Assay Characteristics
» Homogeneous: Add and mix protocols simplify automation for quicker assays without the need for washing, centrifugation or filtering.
» Scaleable: Adaptable protocols in 96-, 384- and 1536-well applications
» Convenient: Use existing liquid handling either in batch or HTS mode with existing FP readers.
Product Overview
