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HTRF cAMP Gi Detection Kit, 1,000 Assay Points
HTRF cAMP Gi Detection Kit, 1,000 Assay Points
Generic HTRF cAMP Gi assay image
| Feature | Specification |
|---|---|
| Application | Second Messenger Detection |
| Sample Volume | 9 µL |
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
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).
HTRF cAMP Gi Detection Kit, 1,000 Assay Points
Generic HTRF cAMP Gi assay image
HTRF cAMP Gi Detection Kit, 1,000 Assay Points
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.
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.
| cAMP Gi | cAMP Gs dynamic | |||
|---|---|---|---|---|
| DOR Antagonists | Naltrindole | Naloxone | Naltrindole | Naloxone |
| Optimal Cells density / well | 6000 | 6000 | 12000 | 12000 |
| S/B | 4 | 4.2 | 2.3 | 2.3 |
| IC50 (nM) | 1.4 | 868 | 2.4 | 934 |
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 |
1,000 assay points
|
Video gallery
HTRF cAMP Gi Detection Kit, 1,000 Assay Points
HTRF cAMP Gi Detection Kit, 1,000 Assay Points
Citations
Resources
Are you looking for resources, click on the resource type to explore further.
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