CIS Bio International
BP 32, Gif/S/Yvette
Paris, 91192
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cAMP Assay Development Kit
cAMP is one of the most established cell transduction signal elements, and its assessment may be the more appropriate way of investigating molecular events occurring at the membrane (e.g. GPCR stimulation). The HTRF cAMP assay is a homogeneous single-step competitive immunoassay based on the interaction between europium cryptate-labeled, anti-cAMP antibody and cAMP-XL665 conjugates. Two different versions of the kit are available: cAMP femtomolar and cAMP dynamic. Both assays are pre-packaged either for 1,000 tests or for 20,000 tests on a 384 low-volume microplate basis (20 μl final volume). They are highly flexible and can easily be further miniaturized by proportional reagent volume reduction without any alteration in assay performance. The two cAMP kit versions share a common "single plate" protocol with two main steps (cell stimulation followed by detection of cAMP) in which cell lysis and detection steps are simultaneously performed at room temperature after 1 hour. This protocol only involves reagent addition to a single FIA microplate and is particularly appropriate for HTS. The assay is compatible with both suspended and adherent cells, and now cell membranes. Like all other HTRF assays, the patented calculation of the fluorescence ratio (665 nm/620 nm) eliminates possible photophysical interference and allows the assay to be unaffected by usual medium conditions like culture medium, serum, biotin and colored compounds.
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