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.
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