HTRF cAMP Gi Detection Kit, 20,000 Assay Points

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HTRF cAMP Gi Detection Kit, 20,000 Assay Points

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This HTRF kit detects changes of cAMP accumulation in response to Gi coupled GPCR activation or inhibition

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
Sample Volume	9 µL

This HTRF kit detects changes of cAMP accumulation in response to Gi coupled GPCR activation or inhibition

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Product Variants

Unit Size: 1,000 Assay Points

Part #:

62AM9PEB

Unit Size: 20,000 Assay Points

Part #:

62AM9PEC

Unit Size: 100,000 Assay Points

Part #:

62AM9PEJ

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Product information

Overview

Based on HTRF technology, the kit contains all the reagents needed to quantify cAMP, ensuring that you benefit from a complete tool for your needs.

Revvity's HTRF cAMP kits are best suited for measuring cAMP modulations in response to Gi-activation, leveraging an optimized combination of robustness and sensitivity.

Specifications

Other Specifications

Application	Second Messenger Detection
Brand	HTRF
Detection Modality	HTRF
Product Group	Kit
Sample Volume	9 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	20,000 Assay Points

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Citations

View product citations for 62AM9PEC on CiteAb

How it works

Assay principle

cAMP kits are based on a competitive format involving a specific antibody labeled with d2 (acceptor) and cAMP coupled to cryptate (donor). This enables the direct characterization of all types of compounds acting on Gi-coupled receptors in either adherent or suspension cells. Native cAMP produced by cells competes with cryptate -labeled cAMP for binding to monoclonal anti-cAMP.

Assay protocol

The protocol is simple and straightforward, with just two incubation steps: - Cell stimulation by the target compounds - cAMP detection using HTRF reagents

Assay details

Product specifications

Designed with signal performance in mind, the kit offers a tailored standard curve ideally suited to the study of Gi coupled GPCRs.

	S/B	IC10 nM	IC50 nM	IC90 nM
cAMP - Gi	31	0.8	3.0	12

Assay validation

Introduction to product validation

The cAMP Gi kit allows enhanced performances for Gi coupled receptor assessment.

The data below shows the pharmacological validation of the Gi kit for the stably expressing delta opioid receptor cell line. Agonists and antagonists were well characterized, showing potency in agreement with the literature.

Validation on agonist dose response

CHO cells stably expressing the delta opioid receptor (DOR) at 6000 cells/well were treated with the SNC-162, a reference agonist, for 45 min.

SNC-162 was well characterized, displaying the right potency in good correlation with the literature.

DOR Agonist SNC-162	cAMP Gi	cAMP Gs dynamic
Optimal Cell density / well	6000	12000
S/B	4.5	2.3
EC50 (nM)	1.6	1.9

Validation on antagonist dose response

CHO cells stably expressing the delta opioid receptor (DOR) at 6000 cells/well were treated with the SNC-162 agonist for 45 min at the EC90 value (10 nM), which is the optimal concentration to use for the antagonist mode experiment.

Fitting the serial dilutions of the Naltrindole and the Naloxone antagonists enabled IC50 calculation of the compounds (potency), which was in perfect correlation with the literature.