cAMP Gs dynamic kit
Optimal performance when applied to Gs activation or inhibition
IP-One Gq ELISA kit HTRF®
This ELISA detects the accumulation of inositol monophosphate, a metabolite produced following phospholipase C activation and used as a proxy of Gq activation
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Proven ELISA format
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Low false negative
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Highly specific
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Accurate pharmacology
Overview
GPCR Gq stimulation is known to induce phospholipase C (PLC) activation and trigger the Inositol phosphate (IP) cascade.
IP1, a downstream metabolite of IP3, accumulates in cells following Gq receptor activation and is stable in the presence of LiCl, making it an ideal GPCR (Gq) functional assay.
Cisbio has developed a monoclonal antibody-based IP-One ELISA to enable simplified and highly accurate measurement of IP1.
Benefits
- SPECIFICALLY TIED TO GQ SIGNALING
- LOW FALSE NEGATIVES
- ACCURATE PHARMACOLOGICAL PROFILES
Assay principle
IP-One ELISA is a competitive immunoassay which uses IP1-HRP and an anti-IP1 monoclonal antibody. The kit comes with 96-well microplates pre-coated with an anti-mouse antibody. Cells are stimulated in the presence of LiCl, causing the accumulation of IP1 upon receptor activation. The protocol consists of two steps following cell stimulation: addition of ELISA components and addition of TMB, the HRP substrate. The HRP reaction is stopped and the optical density (OD) read at 450nm.
Assay protocol
Specifications
IP1, a downstream metabolite of IP3, accumulates in cells following Gq receptor activation.
It is stable in the presence of LiCl, making it ideal for GPCR (Gq) functional assay.
| Detection limit | 10 nM |
|---|---|
| Dynamic range | 10-3000 nM |
| EC50 | 110 nM |
| Specificity | No cross-reactivity with 50 µM Myo-inositol, PIP2, IP2, IP3, IP4 or PIP3 |
Assay performance
The Cisbio IP-One ELISA is compatible with cell lysates, enabling live cell functional assays.
Cells can be stimulated using a variety of plate formats prior to the IP-One ELISA detection steps.
This figure shows the results of an IP-One ELISA performed on cells stimulated in either a 24- or 96-well format. Cells were lysed and then transferred to the ELISA plate provided in the kit. We used 80,000 cells per well in the 96-well plates and 400,000 cells per well in the 24-well format.
Both cell stimulation formats produced similar results, with low standard deviations.
In collaboration with the Faculty of health and medical sciences Denmark - Scientific Presentations
In collaboration with Novartis - Scientific Presentations
Case Study: HTS of GPR-54 with IP-One Assay
In collaboration with Harvard Center for Neurodegeneration and Repair - Scientific Presentations
HTRF products and supporting scientific content to investifgate GPCRs - Flyers
HTRF assays for Oncology and Inflammation
Signaling in the Immune System - Brochures
Webinar: Advancing GPCR drug discovery through Allosteric Modulation
Featuring a panel of experts - Videos
Webinar: Biased ligands in Drug Discovery: the case of TRV027
Biased ligand concept - Videos
Ultimate guides to master Gq assays
Optimizing IP-One assays has never been easier - Guides
Webinar: High-Throughput Screening of GPCRs for Drug Discovery Applications
Featuring a panel of experts - Videos
What every GPCR scientist should know about IP-One
Benefits of Inositol phosphate assays over calcium flux - Guides
Understanding GPCRs is the key to improved DD
In collaboration with GEN - Guides
Guidelines and recommendations for analyzing GPCR 2nd messenger data
Written by a panel of experts - Guides
Product Insert IP-One ELISA / 72IP1PEA-72IP1PED
72IP1PEA-72IP1PED - Product Insert
All your HTRF assays in one document! - Catalog
A guide to Homogeneous Time Resolved Fluorescence
General principles of HTRF - Guides
How HTRF compares to Western Blot and ELISA
Get the brochure about technology comparison. - Brochures
GPCR in Cardiovascular Research: The Relevance of Camp and IP-1 Second Messenger Assays
See how peer researchers monitor GPCR signaling with HTRF - Application Notes
A comprehensive overview of pharmacology's main ligands - Guides
GPCR compounds identification and pharmacological characterization with GTP Gi binding assay
Characterization of candidate molecules through GTP binding assays - Application Notes
GPCRs: the pathway to Discovery
Complete solutions for GPCR Drug Discovery - Brochures
Safety Data Sheet (ENG-GB) IP-One ELISA / 72IP1PEA
72IP1PEA - Safety Data Sheet
Safety Data Sheet (FRA-FR) IP-One ELISA / 72IP1PEA
72IP1PEA - Safety Data Sheet
Safety Data Sheet (DEU) IP-One ELISA / 72IP1PEA
72IP1PEA - Safety Data Sheet
Safety Data Sheet (ELL) IP-One ELISA / 72IP1PEA
72IP1PEA - Safety Data Sheet
Safety Data Sheet (ITA) IP-One ELISA / 72IP1PEA
72IP1PEA - Safety Data Sheet
Safety Data Sheet (SPA) IP-One ELISA / 72IP1PEA
72IP1PEA - Safety Data Sheet
Safety Data Sheet (ENG-GB) IP-One ELISA / 72IP1PEA
72IP1PEA - Safety Data Sheet
Safety Data Sheet (ENG-US) IP-One ELISA / 72IP1PEA
72IP1PEA - Safety Data Sheet
Safety Data Sheet IP-One ELISA / 72IP1PED
72IP1PED - Safety Data Sheet
Safety Data Sheet (DEU) IP-One ELISA / 72IP1PED
72IP1PED - Safety Data Sheet
Safety Data Sheet (ELL) IP-One ELISA / 72IP1PED
72IP1PED - Safety Data Sheet
Safety Data Sheet (FRA-FR) IP-One ELISA / 72IP1PED
72IP1PED - Safety Data Sheet
Safety Data Sheet (ITA) IP-One ELISA / 72IP1PED
72IP1PED - Safety Data Sheet
Safety Data Sheet (SPA) IP-One ELISA / 72IP1PED
72IP1PED - Safety Data Sheet
Safety Data Sheet (ENG-GB) IP-One ELISA / 72IP1PED
72IP1PED - Safety Data Sheet
Safety Data Sheet (ENG-US) IP-One ELISA / 72IP1PED
72IP1PED - Safety Data Sheet
Plate Reader Requirement
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