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Format
96-well strip plate
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Assay Length
3.5 hours
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Sample Type & Volume Required Per Well
Cell Culture Supernates (100 uL), Cell Lysates (100 uL), EDTA Plasma (100 uL), Saliva (100 uL), Urine (100 uL)
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Sensitivity
3.06 pmol/mL
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Assay Range
2.1 - 500 pmol/mL (Cell Culture Supernates, Cell Lysates, EDTA Plasma, Saliva, Urine)
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Specificity
Measures cyclic GMP levels in various samples
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Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
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Interference
No significant interference observed with available related molecules.
Control Available
QC54, Parameter Immunoassay Control Set 870 for cGMP -
Product Summary
The Parameter cGMP Immunoassay is a 3.5 hour competitive enzyme immunoassay designed to measure cGMP in cell culture supernates, plasma, saliva, urine, and cell lysates.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Lysates | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean | 57 | 72 | 178 | 52 | 67 | 195 |
| Standard Deviation | 2.2 | 4 | 9.5 | 4 | 4.4 | 12.7 |
| CV% | 3.9 | 5.6 | 5.3 | 7.7 | 6.6 | 6.5 |
Cell Culture Supernates, EDTA Plasma, Saliva, Urine | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean | 85.2 | 184 | 265 | 80.7 | 188 | 273 |
| Standard Deviation | 6.9 | 12.5 | 14 | 8 | 16.5 | 22.4 |
| CV% | 8.1 | 6.8 | 5.3 | 9.9 | 8.8 | 8.2 |
Recovery
The recovery of cGMP spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
| Cell Culture Media (n=4) | 100 | 91-108 |
| Cell Lysates (n=1) | 92 | 80-98 |
| EDTA Plasma (n=4) | 91 | 74-105 |
| Saliva (n=4) | 102 | 80-123 |
| Urine (n=4) | 90 | 77-98 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of cGMP were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Background: cGMP
Guanosine 3, 5-cyclic monophosphate (cGMP) is formed via the action of guanylate cyclase on GTP. cGMP is present at levels typically 10-100 fold lower than cAMP in most tissues. A variety of stimuli including acetylcholine, insulin, oxytocin, serotonin, and histamine will cause an increase in cGMP levels. Various stimulators of guanylate cyclase (e.g. vasodilators) as well as peptides that relax smooth muscle also increase cGMP concentrations.
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