Cyclic GMP Direct ELISA Kits – Improved Sensitivity

Research Areas

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Specifications

• Assay Type Competitive ELISA
• Sample Types Cell and Tissue Lysates, Urine, Plasma, Saliva, Tissue Culture Media
• Sensitivity 0.31 pmol/mL (Regular), 0.091 pmol/mL (Acetylated)
• Species Cyclic GMP is identical across species
• Assay Duration 2.5 Hours
• Samples/Plate 38 in Duplicate
• Readout Colorimetric, 450 nm
• Standard Curve
• Description

  Assay Principle: 

  The Cyclic GMP Direct ELISA Kit – Improved Sensitivity quantitatively measures Cyclic GMP (cGMP) in saliva, urine, plasma, cell and tissue lysates, and tissue culture media. The Cyclic GMP Direct ELISA Kit – Improved Sensitivity is a competitive ELISA with a run time of 2.5 hours. Please read the complete kit insert for more information before performing this assay.

  Use our provided cGMP 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-rabbit IgG antibody. Add the cGMP peroxidase conjugate and the cGMP rabbit polyclonal antibody. Then incubate the mixture covered at room temperature, shaking for 2 hours. The immunological reaction occurs between the anti-cGMP antibody, the cGMP antigen in the sample or standard, and the cGMP conjugate. As the cGMP concentration in the sample increases, the bound cGMP-peroxidase conjugate decreases, causing a decrease in signal and vice versa. 

  After the 2-hour incubation, wash away the excess cGMP-peroxidase conjugate and add the TMB substrate. The TMB substrate reacts with the bound cGMP-peroxidase conjugate generating a signal detected by a plate reader at 450nm. Use the intensity and the standard curve to calculate the cGMP 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. The enzyme guanylate cyclase (GC) interacts with GTP to generate intracellular forms of cGMP. Phosphodiesterase hydrolysis degrades intracellular forms of cGMP. 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