• Format
    96-well strip plate
  • Assay Length
    3.5 hours
  • 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)
  • Sensitivity
    3.06 pmol/mL
  • Assay Range
    2.1 - 500 pmol/mL (Cell Culture Supernates, Cell Lysates, EDTA Plasma, Saliva, Urine)
  • Specificity
    Measures cyclic GMP levels in various samples
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
QC54, Parameter Immunoassay Control Set 870 for cGMP - Please Inquire
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 Culture Supernates, EDTA Plasma, Saliva, Urine
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean85.218426580.7188273
Standard Deviation6.912.514816.522.4
CV%8.16.85.39.98.88.2

Cell Lysates
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean57721785267195
Standard Deviation2.249.544.412.7
CV%3.95.65.37.76.66.5

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.
cGMP Parameter Assay Kit
Preparation and Storage
  • 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.
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.
    • Long Name
      Cyclic GMP
    • Alternate Names
      cGMP;
    Related Research Areas
    Assay Procedure
    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.
    1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
    2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

    3. 150 µL Diluent to NSB wells & 100 µL Diluent to B0 wells
    4.   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.

    5. 100 µL Standard, Control, or Sample
    6.   Add 100 µL of Standard, Control, or sample to the remaining wells.

    7. 50 µL Conjugate
    8.   Add 50 µL of Conjugate to each well.

    9. 50 µL Primary Antibody Solution
    10.   Add 50 µL of Primary Antibody Solution to each well (except the NSB wells).
    11.   Cover with a plate sealer, and incubate at room temperature for 3 hours on a horizontal orbital microplate shaker.
    12.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

    13. 200 µL Substrate Solution
    14.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

    15. 50 µL Stop Solution
    16. 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.
    Citations:

    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.

    26 Citations: Showing 1 - 10
    Filter your results:

    Species
    Sample Type
    1. Does hypoxia play a role in the development of sarcopenia in humans? Mechanistic insights from the Caudwell Xtreme Everest Expedition
      Authors: L Wandrag, M Siervo, HL Riley, M Khosravi, BO Fernandez, CA Leckstrom, DS Martin, K Mitchell, DZH Levett, HE Montgomery, MG Mythen, MA Stroud, MPW Grocott, M Feelisch, Caudwell X
      Redox Biol, 2017;13(0):60-68.
      Species: Human
      Sample Type: Plasma
    2. REM sleep deprivation induces endothelial dysfunction and hypertension in middle-aged rats: Roles of the eNOS/NO/cGMP pathway and supplementation with L-arginine
      Authors: J Jiang, Z Gan, Y Li, W Zhao, H Li, JP Zheng, Y Ke
      PLoS ONE, 2017;12(8):e0182746.
      Species: Rat
      Sample Type: Tissue Homogenates
    3. Angiotensin-(1-7) Downregulates Diabetes-Induced cGMP Phosphodiesterase Activation in Rat Corpus Cavernosum
      Authors: GS Dhaunsi, M Yousif, B Makki, S Akhtar, IF Benter
      Biomed Res Int, 2017;2017(0):5084961.
      Species: Rat
      Sample Type: Tissue Homogenates
    4. In vitro and in vivo investigation of natural compounds from seed extract of Mucuna pruriens lacking l-DOPA for the treatment of erectile dysfunction
      Authors: N Duangnin, T Phitak, P Pothacharo, P Kongtawele
      Asian Pac J Trop Med, 2017;10(3):238-252.
      Species: Human
      Sample Type: Cell Lysates
    5. Critical role of vascular peroxidase 1 in regulating endothelial nitric oxide synthase
      Authors: Z Liu, Y Liu, Q Xu, H Peng, Y Tang, T Yang, Z Yu, G Cheng, G Zhang, R Shi
      Redox Biol, 2017;12(0):226-232.
      Species: Human
      Sample Type: Cell Lysates
    6. Epigallocatechin-3-gallate ameliorates erectile function in aged rats via regulation of PRMT1/DDAH/ADMA/NOS metabolism pathway
      Authors: D Chen, KQ Zhang, B Li, DQ Sun, H Zhang, Q Fu
      Asian J Androl, 2016;0(0):.
      Species: Rat
      Sample Type: Tissue Homogenates
    7. Lycopene Ameliorates Transplant Arteriosclerosis in Vascular Allograft Transplantation by Regulating the NO/cGMP Pathways and Rho-Associated Kinases Expression
      Oxid Med Cell Longev, 2016;2016(0):3128280.
      Species: Rat
      Sample Type: Plasma
    8. Unbalanced Oxidant-Antioxidant Status: A Potential Therapeutic Target for Coronary Chronic Total Occlusion in Very Old Patients
      Oxid Med Cell Longev, 2016;2016(0):4910829.
      Species: Rat
      Sample Type: Tissue Homogenates
    9. Asymmetric Dimethylarginine Limits the Efficacy of Simvastatin Activating Endothelial Nitric Oxide Synthase
      Authors: CP Hsu, JF Zhao, SJ Lin, SK Shyue, BC Guo, TM Lu, TS Lee
      J Am Heart Assoc, 2016;5(4):.
      Species: Human
      Sample Type: Cell Culture Supernates
    10. Impaired sodium dependent adaptation of arterial stiffness in formerly preeclamptic women: The RETAP - vascular study
      Authors: AM van der Gr, ND Paauw, TJ Toering, M Feelisch, MM Faas, TR Sutton, MM Minnion, JD Lefrandt, SA Scherjon, A Franx, G Navis, AT Lely
      Am J Physiol Heart Circ Physiol, 2016;0(0):ajpheart.0001.
      Species: Human
      Sample Type: Plasma
    11. S-palmitoylation of a Novel Site in the ?2-adrenergic Receptor Associated with a Novel Intracellular Itinerary
      J Biol Chem, 2016;0(0):.
      Species: Human
      Sample Type: Cell Lysates
    12. The effects of phosphodiesterase-5 inhibitor sildenafil against post-resuscitation myocardial and intestinal microcirculatory dysfunction by attenuating apoptosis and regulating microRNAs expression: essential role of nitric oxide syntheses signaling.
      Authors: Zhang Q, Wang G, Yuan W, Wu J, Wang M, Li C
      J Transl Med, 2015;13(0):177.
      Species: Porcine
      Sample Type: Tissue Homogenates
    13. Telmisartan activates endothelial nitric oxide synthase via Ser1177 phosphorylation in vascular endothelial cells.
      Authors: Myojo, Masahiro, Nagata, Daisuke, Fujita, Daishi, Kiyosue, Arihiro, Takahashi, Masao, Satonaka, Hiroshi, Morishita, Yoshiyuk, Akimoto, Tetsu, Nagai, Ryozo, Komuro, Issei, Hirata, Yasunobu
      PLoS ONE, 2014;9(5):e96948.
      Species: Human
      Sample Type: Cell Lysates
    14. Resolvin D1 stimulates alveolar fluid clearance through alveolar epithelial sodium channel, Na,K-ATPase via ALX/cAMP/PI3K pathway in lipopolysaccharide-induced acute lung injury.
      Authors: Wang Q, Zheng X, Cheng Y, Zhang Y, Wen H, Tao Z, Li H, Hao Y, Gao Y, Yang L, Smith F, Huang C, Jin S
      J Immunol, 2014;192(8):3765-77.
      Species: Rat
      Sample Type: Cell Lysates
    15. Simultaneous exposure to Escherichia coli heat-labile and heat-stable enterotoxins increases fluid secretion and alters cyclic nucleotide and cytokine production by intestinal epithelial cells.
      Authors: Read L, Hahn R, Thompson C, Bauer D, Norton E, Clements J
      Infect Immun, 2014;82(12):5308-16.
      Species: Human
      Sample Type: Cell Lysates
    16. MicroRNA-378 controls classical brown fat expansion to counteract obesity.
      Authors: Pan, Dongning, Mao, Chunxiao, Quattrochi, Brian, Friedline, Randall, Zhu, Lihua J, Jung, Dae Youn, Kim, Jason K, Lewis, Brian, Wang, Yong-Xu
      Nat Commun, 2014;5(0):4725.
      Species: Mouse
      Sample Type: Tissue Homogenates
    17. Critical role of exogenous nitric oxide in ROCK activity in vascular smooth muscle cells.
      Authors: Maruhashi, Tatsuya, Noma, Kensuke, Iwamoto, Yumiko, Iwamoto, Akimichi, Oda, Nozomu, Kajikawa, Masato, Matsumoto, Takeshi, Hidaka, Takayuki, Kihara, Yasuki, Chayama, Kazuaki, Nakashima, Ayumu, Goto, Chikara, Liao, James K, Higashi, Yukihito
      PLoS ONE, 2014;9(10):e109017.
    18. Oxygen mediates vascular smooth muscle relaxation in hypoxia.
      Authors: Dada, Jessica, Pinder, Andrew G, Lang, Derek, James, Philip E
      PLoS ONE, 2013;8(2):e57162.
      Species: N/A
      Sample Type: Protein
    19. Decompensated heart failure is associated with reduced corin levels and decreased cleavage of pro-atrial natriuretic peptide.
      Authors: Ibebuogu UN, Gladysheva IP, Houng AK
      Circ Heart Fail, 2011;4(2):114-20.
      Species: Human
      Sample Type: Plasma
    20. The role of interleukin-6 in UVA protection against UVB-induced immunosuppression.
      Authors: Reeve VE, Tyrrell RM, Allanson M, Domanski D, Blyth L
      J. Invest. Dermatol., 2009;129(6):1539-46.
      Species: Mouse
      Sample Type: Tissue Homogenates
    21. Thiophenecarboxylate suppressor of cyclic nucleotides discovered in a small-molecule screen blocks toxin-induced intestinal fluid secretion.
      Authors: Tradtrantip L, Yangthara B, Padmawar P, Morrison C, Verkman AS
      Mol. Pharmacol., 2009;75(1):134-42.
      Species: Hamster
      Sample Type: Cell Lysates
    22. Hematologic, biochemical, and cardiopulmonary effects of L-arginine supplementation or phosphodiesterase 5 inhibition in patients with sickle cell disease who are on hydroxyurea therapy.
      Authors: Little JA, Hauser KP, Martyr SE, Harris A, Maric I, Morris CR, Suh JH, Taylor J, Castro O, Machado R, Kato G, Gladwin MT
      Eur. J. Haematol., 2009;82(4):315-21.
      Species: Human
      Sample Type: Plasma
    23. Decreased nitric oxide products in the urine of patients undergoing cardiac surgery.
      Authors: Lema G, Urzua J, Jalil R, Canessa R, Vogel A, Moran S, Fajuri A, Carvajal C, Aeschlimann N, Jaque MP
      J. Cardiothorac. Vasc. Anesth., 2009;23(2):188-94.
      Species: Human
      Sample Type: Urine
    24. Terfenadine-induced apoptosis in human melanoma cells is mediated through Ca2+ homeostasis modulation and tyrosine kinase activity, independently of H1 histamine receptors.
      Authors: Jangi SM, Ruiz-Larrea MB, Nicolau-Galmes F, Andollo N, Arroyo-Berdugo Y, Ortega-Martinez I, Diaz-Perez JL, Boyano MD
      Carcinogenesis, 2008;29(3):500-9.
      Species: Human
      Sample Type: Cell Lysates
    25. cGMP-independent anti-tumour actions of the inhibitor of soluble guanylyl cyclase, ODQ, in prostate cancer cell lines.
      Authors: Haramis G, Zhou Z, Pyriochou A, Koutsilieris M, Roussos C, Papapetropoulos A
      Br. J. Pharmacol., 2008;155(6):804-13.
      Species: Human
      Sample Type: Cell Culture Supernates
    26. Successive action of meprin A and neprilysin catabolizes B-type natriuretic peptide.
      Authors: Pankow K, Wang Y, Gembardt F, Krause E, Sun X, Krause G, Schultheiss HP, Siems WE, Walther T
      Circ. Res., 2007;101(9):875-82.
      Species: Human
      Sample Type: Cell Lysates
    Expand to show all 26 Citations
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