Cyclic GMP CLIA Kit (High-Sensitivity), Species Independent

Description

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

Species Reactivity

Wide Species

Applications

Direct CLIA (Chemiluminescent Immunoassay)

Relevant Diseases

Neurological Disorders

Sensitivity

0.034 pmol/ml

Platform

Microplate

Detection Method

Colorimetric Assay

Sample Type

Cell lysates, EDTA Plasma, Saliva, Tissue, Tissue Culture Media, Urine

Assay Type

CLIA Kits

Precision

Intra Assay Precision (Regular): Two human urine samples were diluted with Sample Diluent and run in replicates of 22 in an assay. The mean and precision of the calculated cGMP concentrations were: Sample 1- 0.053 pmol/mL, 10.6% CV Sample 2- 0.225 pmol/mL, 11.8% CV Intra Assay Precision (Acetylated): Two human urine samples were diluted with Sample Diluent, acetylated and run in replicates of twenty-two in an assay. The mean and precision of the calculated cGMP concentrations were: Sample 1- 0.032 pmol/mL, 8.9% CV Sample 2- 0.018 pmol/mL, 8.3% CV Inter Assay Precision (Regular): Two human urine samples were diluted with Sample Diluent and run in duplicates in fourteen assays run over multiple days by four operators. The mean and precision of the calculated cGMP concentrations were: Sample 1- 0.213 pmol/mL, 14.9% CV Sample 2- 0.083 pmol/mL, 18.0% CV Inter Assay Precision (Acetylated): Two human urine sample weres diluted with Sample Diluent, acetylated and run in duplicates in eight assays run over multiple days by four operators. The mean and precision of the calculated cGMP concentrations were: Sample 1- 0.07 pmol/mL,15.1% CV Sample 2- 0.04 pmol/mL, 15.1% CV

Number Of Samples

39 samples in duplicate

Linear Range

Regular: 0.0469 - 3 pmol/ml. Acetylated: 0.03125 - 1 pmol/ml

Components

Coated White 96 Well Plates 1 Each
Cyclic GMP Standard 70 µl
StressXpress® Cyclic GMP CLIA Antibody 3 ml
StressXpress® Cyclic GMP CLIA Conjugate 150 µl
Conjugate Diluent 3 ml
Sample Diluent 28 ml
Plate Primer 5 ml
Acetic Anhydride 2 ml
Triethylamine 4 ml
Wash Buffer Concentrate 30 ml
Substrate Solution A 6 ml
Substrate Solution B 6 ml
Plate Sealer 1 Each

Shipping

Blue Ice

Storage

4°C

Research Use Only

For research use only

Official Name

Cyclic GMP

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