The recovery of cGMP spiked to levels throughout the range of the assay in various matrices was evaluated.
Average % Recovery
Cell Culture Media (n=4)
Cell Lysates (n=1)
EDTA Plasma (n=4)
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
Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.
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.
Refer to the product for complete assay procedure.
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
150 µL Diluent to NSB wells & 100 µL Diluent to B0 wells
Add 150 µL of the appropriate diluent to the non-specific binding (NSB) wells and 100 µL of the appropriate diluent to the zero standard (B0) wells.
100 µL Standard, Control, or Sample
Add 100 µL of Standard, Control, or sample to the remaining wells.
50 µL Conjugate
Add 50 µL of Conjugate to each well.
50 µL Primary Antibody Solution
Add 50 µL of Primary Antibody Solution to each well (except the NSB wells).
Cover with a plate sealer, and incubate at room temperature for 3 hours on a horizontal orbital microplate shaker.
Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
200 µL Substrate Solution
Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
50 µL Stop Solution
Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
R&D Systems personnel manually curate a database that contains references using R&D Systems products.
The data collected includes not only links to publications in PubMed,
but also provides information about sample types, species, and experimental conditions.