Description
Key facts
Detection method:Fluorescent,
Sample types:Urine, Plasma, Tissue Extracts, Cell culture media, Cell Lysate,
Assay type:Competitive,
Sensitivity:= 1 nM,
Assay time:3h,
Assay Platform:Microplate reader
Product details:
cAMP Assay Kit (Competitive ELISA, Fluorometric) ab138880 measures cyclic AMP activity in cell extracts, tissue extracts and other biological fluids with a sensitivity of 1nM.
How the cAMP assay works
The cAMP assay is based on the competition between HRP-labeled cAMP and free cAMP present in the sample for cAMP antibody binding sites.
In the absence of cAMP, HRP-cAMP conjugate is bound to anti-cAMP antibody exclusively. Free cAMP (unlabeled) present in the test sample competes with HRP-cAMP for anti-cAMP antibody, therefore displacing HRP-cAMP and inhibiting its binding to the antibody.
The fluorescence observed (Ex/Em = 540/590 nm) is proportional to the activity of HRP-cAMP conjugate: the more cAMP present in the test samples, the lower the fluorescence detected.
cAMP assay protocol summary
- add standards and samples to appropriate wells
- add HRP-cAMP conjugate to samples & standards
- incubate RT 2 hours
- wash wells
- add detector (AbRed Working solution)
- incubate RT 15 – 60 minutes
- measure fluorescence (Ex/Em = 540/590 nm)
How other researchers have used cAMP assay kit ab138880
cAMP Assay Kit ab138880 has been used in a variety of sample types:
- HEK293T cells (1)
- isolated cardiac mitochondria (2)
- sperm samples (3)
References: 1 - Ruiqi Liu et al. 2023; 2 - Maura Greiser et al. 2023; 3 - Qifu He et al. 2023
Related and recommended products
cAMP is commonly used to study signal transduction pathways. Other kits to measure cAMP include:
- cAMP Assay Kit (Competitive ELISA, Fluorometric)
ab323510
, and cAMP ELISA Kit - Intracellular
ab323511
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-Multi, Appropriate long-term storage conditions-Multi, Storage information-Please refer to protocols
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Cyclic adenosine monophosphate (cAMP) serves as an essential secondary messenger in various signal transduction pathways. Known also by its chemical abbreviation cAMP this small molecule has a mass of approximately 329.21 g/mol. cAMP is synthesized from ATP by the enzyme adenylate cyclase which is present in many cell types notably in neurons and various endocrine tissues. cAMP acts by activating protein kinase A (PKA) which then phosphorylates target proteins to alter cellular function.
Biological function summary
CAMP regulates numerous cellular processes including metabolism gene expression and cell growth. It functions as an important part of G protein-coupled receptor (GPCR) signaling pathways where its concentration can significantly change influenced by external signals. cAMP modulates the activity of ion channels and various enzymes often participating in feedback loops to maintain cellular homeostasis.
Pathways
CAMP integrates into critical signal transduction pathways such as the cyclic AMP-dependent pathway and the calcium signaling pathway. It works closely with proteins like CREB (cAMP response element-binding protein) to regulate gene transcription. cAMP also impacts other molecules like phosphodiesterases which break it down maintaining its transient nature in response to stimuli.
Irregular cAMP levels associate with pathologies like heart failure and certain types of cancer. Disruptions in cAMP signaling can lead to abnormal cell growth or inadequate cellular response to hormonal signals. Through its interaction with proteins like CFTR (cystic fibrosis transmembrane conductance regulator) cAMP influences disorders such as cystic fibrosis highlighting its critical role in maintaining cellular function and signaling balance.
