Overview and Description
HitHunter cAMP Assays are applicable for cell based screening of G-protein coupled GPCR activation in a homogeneous, non-radioactive microtiter plate format. HitHunter cAMP assays use Enzyme Fragment Complementation (EFC) for sensitive detection though signal amplification and fewer false positives.
DiscoveRx offers several different configurations based on different applications, readouts, sample and cell types.
Features and Benefits
» Directly assess either Gi- or Gs-coupled GPCRs in cells
» Large signal to background ratios for superior signal differentiation and assay
» Chemiluminescent detection reduces compound interference, lowering false
» Scaleable protocols for 96-, 384- and 1536-well technologies
» No specialized instruments required
Measuring cAMP using HitHunter EFC detection
HitHunter cAMP Assays are competitive immunoassays in which free cAMP from cell lysates compete for antibody binding against labeled ED-cAMP conjugate. ED is a small peptide fragment of ß-galactosidase (ß-gal). In the absence of free cAMP, ED-cAMP conjugates are captured by the antibody and are unavailable for complementation, resulting in low signal. In the presence of free cAMP, antibody sites are occupied, leaving ED-cAMP conjugate free to complement with EA, forming active ß-gal EFC enzyme for substrate hydrolysis to produce a positive chemiluminescent signal. This positive signal is generated in direct proportion to the amount of free cAMP.
» ED-cAMP conjugate label
» cAMP Antibody
» Lysis Buffer
» EFC detection reagents
» cAMP standard
» Luminometer, Multimode Microplate Reader or CCD Camera
Selecting a cAMP Assay
Several configurations are available for different applications with a variety of requirements for throughput, readout, sample and cell types. Performance and throughput expectations and the quality of the cell type dictate which HitHunter cAMP Assay is optimum for a particular target. Visit the DiscoveRx website for recommendations based on your application needs.
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