Invitrogen™ Cyclic AMP Competitive ELISA Kit

Product Code. 10425545

Brand: Invitrogen™ EMSCAMPL

Description

Competitive ELISA

The cyclic AMP Competitive ELISA quantitates cAMP in cell or tissue lysates. The assay will exclusively recognize both natural and recombinant cyclic AMP.

Principle of the method The cyclic AMP solid-phase sandwich ELISA (enzyme-linked immunosorbent assay) is designed to measure the amount of the target bound between a matched antibody pair. A target-specific antibody has been pre-coated in the wells of the supplied microplate. Samples,standards,or controls are then added into these wells and bind to the immobilized (capture) antibody. The sandwich is formed by the addition of the second (detector) antibody,binding to the target on a different epitope from the capture antibody. A conjugated enzyme has been incorporated into the assay. After incubation and washing steps to rid the microplate of unbound substances,a substrate solution is added that reacts with the enzyme-antibody-target complex to produce measurable signal. The intensity of this signal is inversely proportional to the concentration of target present in the original specimen.

Rigorous validation

Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity,specificity,precision,and lot-to-lot consistency. See manual for more information on validation.

Cyclic adenosine monophosphate (cAMP) plays a key role as an intracellular second messenger for transduction events that follow a number of extracellular signals. The G-Protein Coupled Receptors (GPCR) is the largest family of cell surface receptors. They can be activated by different ligands, such as neurotransmitters, hormones, ions, small molecules, peptides, and other physiological signaling molecules. Typically, the binding of the ligands to its receptor resulting in the activation of G-proteins, in return, activates the effector adenylyl cyclase evoking the production of cAMP. The activation of a protein kinase by cAMP results in the phosphorylation of substrate proteins. Currently successful drugs in marketing have been developed to target these receptors. Among the GPCRs, ∽367 receptors are potential drug development targets, but only about 20 have been used to generate therapeutically and commercially successful drugs so far. Because the involvement of cAMP can amplify the response of the ligand binding, the second messenger cAMP has been largely employed to monitor the activation of the GPCR to facilitate the therapeutic drug discovery.

Specifications

• Assay Range: 0.078-20 pmol/mL
• Assay Sensitivity: 0.037 pmol/mL
• Description: Cyclic AMP Competitive ELISA Kit
• Immunoassay Kit Format: Colorimetric ELISA (p-NPP substrate) with Plates
• Product Type: ELISA Kit
• Sample Type: Cell Lysates, Tissue Homogenates
• Specificity: No significant cross-reaction with AMP, ATP, cGMP, GMP, GTP, cUMP, or CTP
• Sufficient For: 96 microplate wells
• Target Species: Human
• For Use With (Equipment): Colorimetric Microplate Reader
• Interassay CV: 7.1%
• Intraassay CV: 6.5%
• Kit Contents: Pre-coated 96-well plate, Detection Antibody, AP Conjugate, Neutralizing Reagent, Triethylamine, Acetic Anhydride, Standard, 20X Wash Buffer, pNpp Substrate Solution, Stop Solution, Plate Sealer, Plate Sealer, 1 ea
• Label or Dye: AP
• Measurement: Absorbance at 405nm minus absorbance at 570nm
• Precision: Dependent upon sample type, level and assay
• Quantity: 96 Tests
• Regulatory Status: RUO
• Sample Volume: Cell Lysate,100 μL; Tissue Homogenate,100 μL
• Storage Requirements: -20°C
• Target: Human
• Test Time: 1 hr. 20 min.
• Total Assay Time: 3 hr.