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Proteomics & Protien Expression

cAMP Assays

Sigma® has prepared product lists for commonly used protocols from Cold Spring Harbor Protocols.

View the entire protocol on BioSupplyNet.com.

Selected Protocol from "Protein-Protein Interactions", 2nd Edition, E. Golemis, P. Adam.

cAMP Assays

Gouzel Karimova and Daniel Ladant
Unite Postulante de Biochimie des Interactions Macromoleculaires, Departement de Biologie Structurale et Chimie,
CNRS URA 2185, Institut Pasteur, Paris, France


ABSTRACT
cAMP measurements are obtained using an ELISA assay (Harlow and Lane 1988). Commercial radio-immunoassays, or ELISA kits, to assay cAMP can be purchased from various manufacturers. In our laboratory we use the homemade, less-expensive ELISA that is described below. In outline, this assay is based on the ability of soluble cAMP in bacterial extracts to compete with binding of an alkaline phosphatase (AP)-conjugated anti-cAMP antibody to immobilized cAMP. The surface-bound, immobilized anti-cAMP-AP is inversely correlated to the concentration of cAMP in the extract. Known concentrations of soluble cAMP are used to calibrate the assay.

 

 

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Products Available for this Protocol
Protocol Material Description Product #  Product Name Add to Cart
Buffers, Solutions, and Reagents      
BSA A3156 Albumin, essentially globulin-free from bovine serum
0.1 M Na2CO3 (pH 9.5) S7795 Sodium carbonate, SigmaUltra
Dimethyl formamide (DMF) D4551 N,N-Dimethylformamide
N-Ethoxy-carbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) 149837 2-Ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline
50 mM HEPES (pH 7.5) H4034 HEPES, Biotechnology Performance Certified
NaCl S3014 Sodium chloride
0.1% Tween-20 P9416 TWEEN® 20,for molecular biology, viscous liquid
Cells      
Bacterials strains to be assayed for cAMP ECACC cell lines Cell Lines  
Antibodies      
Rabbit anti-cAMP antiserum A0670 Anti-Adenosine 3',5'-Cyclic Monophosphate (cAMP) antibody produced in rabbit, whole antiserum, lyophilized powder

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