cGMP Parameter Assay Kit

  (31 citations)     
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Assay Procedure
Citations (31)
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  • 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
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

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

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 [Biotin]
Product Datasheets

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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.

31 Citations: Showing 1 - 10
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Species
Sample Type
  1. Xyloketal B exerts antihypertensive effect in renovascular hypertensive rats via the NO-sGC-cGMP pathway and calcium signaling
    Authors: LY Zhao, J Li, XQ Huang, GH Wang, XF Lv, WF Meng, WL Chen, JY Pang, YC Lin, HS Sun, GL Wang, YM Du
    Acta Pharmacol. Sin., 2018;0(0):.
    Species: Rat
    Sample Type: Cell Lysates
  2. Inhibition of Phosphodiesterase 5 and Increasing the Level of Cyclic Guanosine 3',5' Monophosphate by Hydroalcoholic Achillea wilhelmsii C. Koch Extract in Human Breast Cancer Cell Lines MCF-7 and MDA-Mb-468
    Authors: R Saravani, HR Galavi, A Shahraki
    Breast Cancer (Auckl), 2017;11(0):1178223417690.
    Species: Human
    Sample Type: Cell Lysates
  3. Novel H2S-NO hybrid molecule (ZYZ-803) promoted synergistic effects against heart failure
    Authors: D Wu, Q Hu, Y Xiong, D Zhu, Y Mao, YZ Zhu
    Redox Biol, 2017;15(0):243-252.
    Species: Mouse
    Sample Type: Plasma
  4. Carbon monoxide prevents TNF-?-induced eNOS downregulation by inhibiting NF-?B-responsive miR-155-5p biogenesis
    Authors: S Choi, J Kim, JH Kim, DK Lee, W Park, M Park, S Kim, JY Hwang, MH Won, YK Choi, S Ryoo, KS Ha, YG Kwon, YM Kim
    Exp. Mol. Med., 2017;49(11):e403.
    Species: Human
    Sample Type: Cell Lysates
  5. 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
  6. 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
  7. 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
  8. 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
  9. 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
  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. Alternative hematological and vascular adaptive responses to high-altitude hypoxia in East African Highlanders
    Authors: Hoi I Cheong
    Am. J. Physiol. Lung Cell Mol. Physiol, 2016;0(0):ajplung.00451.
    Species: Human
    Sample Type: Urine
  12. 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
  13. 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
  14. 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
  15. 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
  16. 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
  17. 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
  18. 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
  19. 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
  20. 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
  21. 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.
  22. 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
  23. 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
  24. 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
  25. 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
  26. 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
  27. 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
  28. 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
  29. 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
  30. 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
  31. 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
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