Measurement of cAMP for Gαs- and Gαi Protein-Coupled …
Nov 20, 2017 · The Cisbio cAMP assay uses homogeneous time-resolved fluorescence technology (HTRF) to measure cAMP in a non-separation, high throughput format. These kits are based on a competitive immunoassay using Eu 3+ cryptate-labeled anti-cAMP antibody and d2-labeled cAMP ( Figure 1 ).
cAMP Assay - an overview | ScienceDirect Topics
Fluorescence polarization (FP)-based cAMP measurement kits are now commercially available from Perkin Elmer, 592,593 Molecular Devices, 594 and Amersham (GE Healthcare). In this assay, as endogenous cAMP increases, it competes for trace fluorescent-labeled cAMP binding to a cAMP antibody.
GloSensor™ cAMP Assay Protocol - Promega Corporation
The GloSensor™ cAMP Assay provides an extremely sensitive and easy-to-use format for the interrogation of overexpressed or endogenous GPCRs that signal via changes in the intracellular concentration of cAMP.
Measuring cAMP Levels and Cytotoxicity in a Single Plate Well
Here we describe the multiplexing of a fluorescent real-time cytotoxicity assay with a bioluminescent cAMP assay to study changes in cAMP levels in the context of cytotoxic effects. cAMP is important as a second messenger in a variety of signaling pathways.
A bioluminescent and homogeneous assay for monitoring GPCR
Feb 23, 2024 · In this study, we show cAMP Lumit assay as a highly specific homogeneous bioluminescent assay suitable for high throughput screenings with a large assay window and a wide dynamic range for...
cAMP assays in GPCR drug discovery - PubMed
This review article presents thoughts on the potential of GPCR desensitization assay/cAMP assay, a highly sensitive and versatile platform for high-throughput assays for elucidating novel functions of many orphan GPCRs.
Table 1. [Comparison of popular cAMP assay technologies]. - Assay ...
A luciferase biosensor-based assay that uses genetically encoded biosensor with cAMP binding domain fused to a luciferase. Binding of cAMP to the cAMP binding domain changes the conformation of the biosensor, leading to the activation of luciferase.
cAMP assays in GPCR drug discovery - ScienceDirect
Jan 1, 2017 · This review article presents thoughts on the potential of GPCR desensitization assay/cAMP assay, a highly sensitive and versatile platform for high-throughput assays for elucidating novel functions of many orphan GPCRs.
cAMP detection methods in HTS: selecting the best from the …
Feb 1, 2004 · Two main types of technology for detecting changes in cAMP levels are discussed: accumulation assays and reporter-gene assays. In accumulation assays, changes in intracellular cAMP...
Measurement of cAMP for Gαs- and Gαi Protein-Coupled ... - PubMed
Nov 20, 2017 · The advantages and disadvantages of the various cAMP assay technologies are given in Table 1. This chapter focuses on the Cisbio HTRF cAMP kit to illustrate the methodology and considerations for development of assays that measure intracellular cAMP.
Live-cell imaging of cAMP dynamics | Nature Methods
Dec 28, 2007 · In this review, we examine cAMP biosensors that are currently available for measuring cAMP at the single-cell level, compare their various operating principles and discuss their applications.
The GloSensor™ cAMP Assay provides an extremely sensitive and easy-to-use format for the interrogation of overexpressed or endogenous GPCRs that signal via changes in the intracellular concentration of cAMP.
Cell preparation: a key step for successful cAMP assays.
Jun 11, 2024 · To simplify your life a little, we have compiled all the information you’ll need for successful cAMP assays in 4 different guides. Get yours now! For research use only.
cAMP Assay - an overview | ScienceDirect Topics
In this assay, cAMP is detected by the rabbit antibody to cAMP, linked to a marker enzyme, phosphodiesterase. The assay components react for several hours and the plate is washed and a substrate for the detection enzyme added.
Luciferase-based GloSensor™ cAMP assay: Temperature
Here, we describe the temperature sensitivity and reversibility of the GloSensor™ cAMP assay, and which GloSensor™ version is optimal for measuring cytosolic cAMP. We also present a detailed protocol for monitoring cAMP levels in live cells …
Selecting a cyclic AMP kit for assaying GPCR target activation
How to select a cAMP assay. In a cell-based assay, the intracellular cAMP level depends not only on the agonist or antagonist activity but also on the cellular model used.
cAMP-Glo™ Assay - Promega
Powered by Ultra-Glo™ Recombinant Luciferase, the bioluminescence-based cAMP-Glo™ assay provides a safe alternative to radioactive methods, and avoids the interference issues that can be problematic for fluorescence-based methods.
cAMP assays - PubMed
Apr 1, 2007 · This assay is based on the ability of soluble cAMP in bacterial extracts to compete with binding of an alkaline phosphatase (AP)-conjugated anti-cAMP antibody to immobilized cAMP. The surface-bound, immobilized anti-cAMP-AP is inversely correlated to the concentration of cAMP in the extract.
Measurement of cAMP in an Undergraduate Teaching Laboratory …
Feb 14, 2012 · The Perkin Elmer assay system called ALPHAscreen (Amplified Luminescent Proximity Homogenous Assay) enables rapid, high-throughput detection of proteins, such as phosphorylated kinases , and other signal transduction molecules, such as cAMP .
cAMP binding protein assay for widespread use in cell
cAMP plays a critical role in intracellular signaling pathways that regulate proliferation or differentiation. The cAMP binding protein assay, using a naturally derived cAMP binding protein, is one of the most widely used methods for cAMP determination.