Cyclic GMP High-Sensitivity CLIA

$590.00

The Cyclic GMP CLIA Kit (High-Sensitivity) is designed to quantitatively measure cGMP present in lysed cells, EDTA plasma, urine, saliva and tissue culture media samples.

SKU: SKT-210-96 Categories: , ,

Cyclic GMP High-Sensitivity CLIA

The Cyclic GMP High-Sensitivity CLIA is For Research Use Only

Size: 1×96 wells
Sensitivity: 0.034 pmol/ml
Dynamic Range: Regular: 0.0469 – 3 pmol/ml. Acetylated: 0.03125 – 1 pmol/ml
Incubation Time: 5 minutes
Sample Type: Cell lysates, EDTA Plasma, Saliva, Tissue, Tissue Culture Media, Urine
Sample Size: 50 μL

Product manufactured in Canada by StressMarq.


Assay Principle

For samples where the levels of cGMP are expected to be relatively high, the regular format for the assay can be used. For samples with expected low levels of cGMP, an optional acetylation protocol can be used. The kit is unique in that all samples and standards are diluted into an acidic Sample Diluent, which contains special additives and stabilizers, for cGMP measurement. This allows plasma, urine and saliva samples to be read in an identical manner to lysed cells. Acidified samples of cGMP are stable and endogenous phosphodiesterases are inactivated in the Sample Diluent. A cGMP standard is provided to generate a standard curve for the assay and all samples should be read off the standard curve. A white microtiter plate coated with an antibody to capture mouse IgG is provided and a neutralizing Plate Primer solution is added to all the used wells. Standards or diluted samples, either with or without acetylation, are pipetted into the primed wells. A cGMP-peroxidase conjugate is added to the standards and samples in the wells. The binding reaction is initiated by the addition of a mouse monoclonal antibody to cGMP to each well. After an overnight incubation at 4°C, the plate is washed and the chemiluminescent substrate is added. The substrate reacts with the bound cGMP-peroxidase conjugate to produce light The generated light is detected in a microtiter plate reader capable of reading luminescence. The concentration of the cGMP in the sample is calculated, after making suitable correction for the dilution of the sample, using software available with most plate readers.


Related Products

Cyclic GMP EIA Kit
Cyclic AMP High-Sensitivity CLIA
Cyclic AMP EIA Kit

Additional Information

Assay Background


Guanosine 3’, 5’-cyclic monophosphate (cyclic GMP; cGMP) is a critical and multifunctional second messenger present at levels typically 10-100 fold lower than cAMP in most tissues. Intracellular cGMP is formed by the action of the enzyme guanylate cyclase on GTP and degraded through phosphodiesterase hydrolysis (1-3). Guanylate cyclases (GC) are either soluble or membrane bound (3,4). Soluble GCs are nitric oxide responsive, whereas membrane bound GCs respond to hormones such as acetylcholine, insulin and oxytocin. Other chemicals like serotonin and histamine also cause an increase in cGMP levels (5,6). Cyclic GMP regulates cellular composition through cGMP-dependent kinase, cGMP-dependent ion channels or transporters, and through its hydrolytic degradation by phosphodiesterase (1,7)

Typical Standard Curve


Cyclic GMP High-Sensitivity CLIA

Manual

Product Manual