Adenosine 3’, 5’ cyclic monophosphate (cAMP) is an important second messenger in intracellular signal transduction. Monitoring cAMP levels is one of the most common ways to screen for agonists and antagonists of GPCRs. ACTOne™ cAMP Fluorimetric ELISA Assay Kit is based on the competition between HRP-labeled cAMP and free cAMP for a fixed number of cAMP antibody binding sites. In the absence of free cAMP, HRP-cAMP conjugate is bound to anti-cAMP antibody exclusively. In the presence of free cAMP, HRP-cAMP is displaced from the HRP-cAMP/anti-cAMP antibody complex.
ACTOne™ cAMP Florimetric ELISA Assay Kit provides the sensitive method for detecting cAMP level in biochemical or cell-based assay system. Compared to other ELISA cAMP assay kits, our kit eliminates the tedious acetylation step. The kit uses Amplite™ Red as a fluorimetric substrate to quantify the HRP activity. The assay can be performed in a convenient 96-well or 384- well microtiter-plate format and easily adapted to automation. The fluorescent product formed is proportional to the activity of HRPcAMP conjugate.
|Component A: cAMP Standard||1 vial (33 µg)|
|Component B: Assay Buffer||500 mL|
|Component C: HRP-cAMP Conjugate||1 vial|
|Component D: 10X Wash Solution||2x100 mL|
|Component E: Cell Lysis Buffer||100 mL|
|Component F: 3% H2O2||50 µL|
|Component G: Amplite™ Red||1 vial|
|Component H: Anti-cAMP Ab Coated 96-Well Plate||10 plates|
|Component I: Substrate Buffer||2x100 mL|
Note: Do not freeze Anti-cAMP Ab Pre-coated 96-well plate (Component H), store it at 4oC.
|Operating Temperature Range||Room temperature|
|Shipping Condition||Blue ice|
|Storage and shelf-life||4°C, 6 months|