cAMP Assay Kit uses a sensitive colorimetric competitive ELISA method to quantify cAMP (cyclic AMP) levels in samples from mammals and other species. It has a sensitivity of 0.02 µM.The cAMP assay protocol uses a 96-well plate supplied coated with Protein G. The Protein G binds an anti-cAMP antibody which is added to the plate.HRP-labeled cAMP, and free cAMP within a sample, are then added to each well of the plate. The cAMP assay is based on the competition between the HRP-labeled cAMP and the free cAMP for the fixed number of cAMP antibody binding sites on the plate.The more free cAMP is present, the less HRP-labeled cAMP is bound. After a washing step, the amount of bound HRP-labeled cAMP is detected using a colorimetric HRP substrate measured by absorbance at OD 450 nm. The level of signal is compared to a standard curve for free cAMP, which is run at the same time as the samples.This kit uses an acetylation step to achieve the maximum sensitivity.
|Sample type||Urine, Serum, Other biological fluids, Tissue Extracts, Cell Lysate, Cell culture media|
|Sensitivity||> 0.02 µM|
|Range||0.02 µM – 2 µM|
|Assay time||3h 30m|
|Species reactivity||Reacts with: Mammals, Other species|
cAMP Assay Protocol Summary:
– add prepared samples and standards to tubes
– add neutralizing buffer
– add acetylating reagent mix, vortex and incubate for 10 min at room temp
– add assay buffer
– transfer to protein-G 96-well plate
– add cAMP antibody and incubate for 1 hr at room temp
– add cAMP-HRP and incubate for 1 hr at room temp
– wash with assay buffer
– add HRP developer and incubate for 1 hr at room temp
– stop reaction with HCl
– analyze with microplate reader
Store kit at -20°C immediately on receipt and check below for storage for individual components. Kit can be stored for 1 year from receipt, if components have not been reconstituted. Reconstituted components are stable for 1-2 months.Aliquot components in working volumes before storing at the recommended temperature.
Avoid repeated freeze-thaws of reagents.
100tests ; 200tests ; 500tests
Celltechgen provides high-quality reagents and materials for research use only. For proper handling of potentially hazardous chemicals, please request the Safety Data Sheet (SDS) provided for the product.