Bioluminescent Detection of cAMP to Monitor GPCR Activity
The cAMP-Glo™ Max Assay is a homogeneous, bioluminescent, high-throughput assay to measure cyclic AMP (cAMP) levels in cells. Compounds that modulate GPCRs coupled with adenylate cyclase typically alter intracellular cAMP levels. The assay monitors cAMP levels in cells in response to the effect of agonists, antagonists or test compounds on G protein-coupled receptors (GPCRs), and is based on the principle that cyclic AMP (cAMP) stimulates protein kinase A (PKA) holoenzyme activity, decreasing available ATP and leading to decreased light production in a coupled luciferase reaction.
Improved Format for Better Performance
This improved version of the cAMP-Glo™ Assay combines the lysis and cAMP reaction buffers into the cAMP-Glo™ ONE Buffer, streamlining the protocol and speeding up the assay. The ONE Buffer is supplied at 5X concentration, which provides increased flexibility for starting cell culture volumes.
The cAMP-Glo™ Max Assay allows easy quantitation of cAMP levels in broad range of cell types, and can be performed in 96-, 384- or 1536-well plates. Cells are induced with a test compound for an appropriate period of time to modulate cAMP levels. After induction, cells are lysed and the cAMP released stimulates protein kinase A in the reagent. The Kinase-Glo® Reagent is then added to terminate the PKA reaction and detect the remaining ATP via a luciferase reaction. Plates are read using a microplate-reading luminometer. The half-life for the luminescent signal is greater than 4 hours, providing ample time to read plates and eliminating the need for luminometers with reagent injectors.
Cat.# V1681 contains sufficient reagents for 2 plates (96- or 384-well format).
Cat.# V1682 contains sufficient reagents for 20 plates (96- or 384-well format).
Cat.# V1683 contains sufficient reagents for 200 plates (96- or 384-well format).
cAMP production in cells and the cAMP-Glo™ Max Assay.
Schematic diagram of cAMP production in cells and the cAMP-Glo™ Max Assay.
Fast and Easy to Use
- Improved—Lysis and cAMP detection steps combined (cAMP-Glo™ ONE Buffer).
- ONE Buffer—5X concentration provides better flexibility for starting cell culture volumes.
- Assay complete in approximately 30 minutes.
Excellent Signal:Noise Ratios
- Best signal:background ratio of all the cAMP assays.
- Signal:Background >200 (with cAMP), >15 (on cells).
Proven Luminescence Technology
- Powered by Ultra-Glo™ Recombinant Luciferase.
- No interference by fluorescent compounds.
- Non-radioactive.
Protocols
Complete Protocol
Specifications
Catalog Number:
Contenido
| Item | Part # | Presentación |
|---|---|---|
|
Protein Kinase A |
V135A | 1 × 20μl |
|
cAMP-Glo™ ONE Buffer |
V166A | 1 × 2.5ml |
|
MgCl2, 1M |
V330A | 1 × 1ml |
|
cAMP, 1mM |
V642B | 1 × 100μl |
|
Kinase-Glo® Substrate |
V672A | 1 × 1 vial |
|
Kinase-Glo® Buffer |
V673A | 1 × 10ml |
SDS
Search for SDSCertificado de Análisis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Condiciones de Almacenaje
U.S. Pat. No. 7,700,310 and other patents and patents pending.
Contenido
| Item | Part # | Presentación | Concentración |
|---|---|---|---|
|
Protein Kinase A |
V135B | 1 × 200μl | |
|
cAMP-Glo™ ONE Buffer |
V166B | 1 × 20ml | |
|
MgCl2 |
V330B | 1 × 5ml | 1M |
|
cAMP, 1mM |
V642A | 1 × 500μl | |
|
Kinase-Glo® Substrate |
V672B | 1 × 1 vial | |
|
Kinase-Glo® Buffer |
V673B | 1 × 100ml |
SDS
Search for SDSCertificado de Análisis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Condiciones de Almacenaje
U.S. Pat. No. 7,700,310 and other patents and patents pending.
Contenido
| Item | Part # | Presentación | Concentración |
|---|---|---|---|
|
Protein Kinase A |
V135B | 10 × 200μl | |
|
cAMP-Glo™ ONE Buffer |
V166B | 10 × 20ml | |
|
MgCl2 |
V330B | 4 × 5ml | 1M |
|
cAMP, 1mM |
V642A | 6 × 500μl | |
|
Kinase-Glo® Substrate |
V672B | 10 × 1 vial | |
|
Kinase-Glo® Buffer |
V673B | 10 × 100ml |
SDS
Search for SDSCertificado de Análisis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Condiciones de Almacenaje
U.S. Pat. No. 7,700,310 and other patents and patents pending.
Resources
No related resources available
Related Products
Productos Similares
GloSensor™ Technology
Biosensores de células vivas para AMPc, GMPc o actividad de proteasas.
E1290, E1291, E2301, E1171
cAMP
Activador de la subunidad catalítica de la proteína quinasa dependiente de AMPc.
V6421
cAMP-Glo™ Assay
Ensayo de alto rendimiento de 45 minutos para medir AMPc en células.
V1501, V1502
AMP-Glo™ Assay
Detección luminiscente de AMP en reacciones bioquímicas.
V5011, V5012
Usado con frecuencia junto con
Kinase-Glo® Luminescent Kinase Assays
Determina la actividad de quinasas purificadas cuantificando el ATP restante tras una reacción quinasa.
V6711, V6712, V6713, V6714, V6071, V6072, V6073, V6074, V3771, V3772, V3773, V3774
NanoBiT® β-Arrestin Recruitment Starter Kit
Live-cell luminescence assay for GPCR signaling and biased agonist profiling.
CS1603B404, CS1603B399
GloMax® Discover System
Lector de microplacas de alto rendimiento para la detección de luminiscencia, fluorescencia y absorbancia.
GM3000
GloMax® Explorer System
Sistema de detección multimodo, flexible y fácil de usar.
GM3500, GM3510