HTRF IP-One Detection Kit, 96 Assay Points

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HTRF IP-One Detection Kit, 96 Assay Points

This ELISA detects the accumulation of inositol monophosphate, a metabolite produced following phospholipase C activation and used as a proxy of Gq activation

For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption and disposal requirements under European REACH regulations (EC 1907/2006).

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Feature	Specification
Application	Second Messenger Detection
Dynamic Range	10 - 3,000 nM
Limit of Detection	10 nM
Sample Volume	50 µL

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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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Product Variants

Unit Size: 96 Assay Points

Part #:

72IP1PEA

List Price

USD 1,147.00

Unit Size: 480 Assay Points

Part #:

72IP1PED

List Price

USD 3,954.00

Product information

Overview

GPCR Gq stimulation is known to induce phospholipase C (PLC) activation and trigger Inositol phosphate (IP) cascade.

IP1, a downstream metabolite of IP3, accumulates in cells following Gq receptor activation and is stable in presence of LiCl, making it an ideal GPCR (Gq) functional assay.

Revvity has developed a monoclonal antibody-based IP-One ELISA to enable simplified and highly accurate measurement of IP1.

Specifications

Other Specifications

Application	Second Messenger Detection
Brand	HTRF
Detection Modality	HTRF
Dynamic Range	10 - 3,000 nM
Limit of Detection	10 nM
Product Group	Kit
Sample Volume	50 µL
Shipping Conditions	Shipped Ambient
Target Class	GPCR
Technology	TR-FRET
Therapeutic Area	Cardiovascular Infectious Diseases Metabolism/Diabetes NASH/Fibrosis Neuroscience Oncology & Inflammation Rare Diseases
Unit Size	96 Assay Points

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Citations

View product citations for 72IP1PEA on CiteAb

How it works

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

Assay details

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.

gpcr-tab3-diagram-gq-coupled-gpcr-activation

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 validation

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.