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Specifications

  • Assay Type Competitive ELISA
  • Sample Types Cell Lysates, Urine, Plasma, Saliva, Tissue Culture Media
  • Sensitivity 0.034 pmol/mL (Regular), 0.023 pmol/mL (Acetylated)
  • Species Cyclic GMP is identical across species
  • Assay Duration 16.5 Hours
  • Samples/Plate 39 in Duplicate (Regular), 40 in Duplicate (Acetylated)
  • Readout Chemiluminescent
  • Standard Curve
  • Description

    Assay Principle: 

    The Cyclic GMP Direct Chemiluminescent ELISA Kit quantitatively measures Cyclic GMP in cell lysates, urine, plasma, saliva, and tissue culture media. The Cyclic GMP Direct Chemiluminescent ELISA Kit is a competitive ELISA with a run time of 16.5 hours. This kit is multi-format and can detect both acetylated and non-acetylated cGMP in a variety of sample types. Please read the complete kit insert for more information before performing this assay.

    Use our provided Cyclic GMP standard to generate a standard curve for the assay. Pipette the standards or diluted samples into a transparent microtiter plate coated with our goat anti-mouse IgG antibody. Add the Cyclic GMP peroxidase conjugate and the Cyclic GMP monoclonal mouse antibody. Then incubate the mixture covered for 16 hours, shaking at 4°C. The immunological reaction occurs between the anti-Cyclic GMP antibody, the Cyclic GMP in the sample or standard, and the Cyclic GMP-peroxidase conjugate. As the Cyclic GMP concentration in the sample increases, the bound Cyclic GMP-peroxidase conjugate decreases, causing a decrease in signal and vice versa. 

    After the 16-hour incubation, wash away the excess Cyclic GMP-peroxidase conjugate and add the chemiluminescent substrate. The chemiluminescent substrate reacts with the bound Cyclic GMP-peroxidase conjugate generating a signal detected by a plate reader. Use the intensity and the standard curve to calculate the Cyclic GMP concentration in the samples.

    Background:

    Guanosine 3’, 5’-cyclic monophosphate (cyclic GMP; cGMP) is a critical and multifunctional second messenger at 10-100 fold lower levels than cAMP in most tissues. Cyclic GMP regulates cellular composition through cGMP-dependent kinase, cGMP-dependent ion channels or transporters, and through its hydrolytic degradation by phosphodiesterase. Intracellular cGMP is formed by the action of the enzyme guanylate cyclase (GC) on GTP and degraded through phosphodiesterase hydrolysis. Guanylate cyclases are either soluble or membrane-bound. Soluble GCs are nitric oxide responsive, whereas membrane-bound GCs respond to acetylcholine, insulin, and oxytocin hormones. Other chemicals like serotonin and histamine also cause an increase in cGMP levels.
  • Structure