Cgmp enzyme immunoassay kit

Manufactured by R&D Systems

The CGMP enzyme immunoassay kit is a laboratory tool used for the quantitative measurement of cyclic guanosine monophosphate (cGMP) levels in various sample types. It provides a standardized and reliable method for detecting and quantifying cGMP, which is an important cellular signaling molecule involved in various physiological processes.

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Lab products found in correlation

Image analysis software, by Media Cybernetics (1 mentions) Ripa buffer, by Thermo Fisher Scientific (1 mentions) Mouse anti actin monoclonal antibody, by Santa Cruz Biotechnology (1 mentions) Mouse monoclonal anti vegf, by Abcam (1 mentions) Ecl cell attachment matrix, by Merck Group (1 mentions) Mouse antibody for α tubulin, by Merck Group (1 mentions) Hydroethidine dhe, by Thermo Fisher Scientific (1 mentions) Enzychrom nadp nadph assay kit, by BioAssay Systems (1 mentions) Griess reagent, by Merck Group (1 mentions) Simvastatin, by Merck Group (1 mentions) 2 7 dichlorofluorescein diacetate dcfh da, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Enzyme immunoassay (50 citations) Enzyme immunoassay kit (44 citations) CGMP immunoassay kit (2 citations)

3 protocols using cgmp enzyme immunoassay kit

Femoral Head Protein Analysis and cGMP/PKG Quantification

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Total protein was isolated by crushing and homogenizing the femoral head with RIPA buffer (Thermo Scientific, USA). Proteins were separated and transferred to membranes following standard protocols and probed using mouse monoclonal anti-VEGF (1:500; Abcam, USA) antibody. Protein expression was visualized on a film with an enhanced chemiluminescence kit (NEN Life Science Products Inc., Boston, MA, USA). Relative protein expression was determined using image analysis software (Media Cybernetics). For the quantitative analysis of protein, the intensity of each test band was expressed as the optical density (OD). β-actin was detected by mouse monoclonal anti-actin antibody (1:1000; Santa Cruz, USA) and was used as an internal control.
Measurement of Tissue cGMP and PKG Activity. Bone tissue cGMP level was measured by enzyme-linked immunosorbent assay using cGMP Enzyme Immunoassay Kit (R&D systems, Minneapolis, MN) according to the manufacturer's directions. PKG activity was measured according to the manufacturer's instructions (Cyclex, Nagano, Japan). Spectrophotometric absorbance was measured at 450 nm, and the results were normalized per mg of protein.

Song Q., Ni J., Jiang H, & Shi Z. (2017). Sildenafil improves blood perfusion in steroid-induced avascular necrosis of femoral head in rabbits via a protein kinase G-dependent mechanism. Acta Orthopaedica et Traumatologica Turcica, 51(5), 398-403.

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Evaluating Endothelial Function in Mice

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Simvastatin, ADMA, SDMA, mouse antibody for α‐tubulin, and Griess reagent were from Sigma‐Aldrich (St Louis, MO). Rabbit antibody for phospho‐eNOS Ser1179 was from Cell Signaling Technology (Beverly, MA). Rabbit antibodies for eNOS and DDAH‐2 were from Santa Cruz Biotechnology (Santa Cruz, CA). The cGMP enzyme immunoassay kit was from R&D Systems (Minneapolis, MN). ECL Cell Attachment Matrix was from Millipore (Bedford, MA). The EnzyChrom NADP⁺/NAD(P)H assay kit was from BioAssay Systems (Hayward, CA). Hydroethidine (DHE) and 2′,7′‐dichlorofluorescein diacetate (DCFH‐DA) were from Molecular Probes (Eugene, OR).

Hsu C., Zhao J., Lin S., Shyue S., Guo B., Lu T, & Lee T. (2016). Asymmetric Dimethylarginine Limits the Efficacy of Simvastatin Activating Endothelial Nitric Oxide Synthase. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 5(4), e003327.

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Quantifying Soluble Guanylate Cyclase Activity

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The cyclase activity reaction was performed
using ∼5 μM full-length sGC (based on the absorbance
at 431 nm) and ∼10 μM catalytic constructs (based on
the absorbance at 280 nm). The reaction assay was conducted in 40
mM HEPES (pH 7.4), 0.5 mM dithiothreitol, 0.3 mM 3-isobutyl-1-methylxanthine,
1 mM GTP, and 3 mM MgCl₂ or MnCl₂. The reaction
mixture was incubated at 15 °C for 20 min and the reaction stopped
with the addition of EDTA (final concentration of 10 mM). The reaction
mixture was stored at −80 °C until cGMP was quantified
using a cGMP enzyme immunoassay kit (R&D) following the manufacturer’s
protocol. Activity was measured in duplicate, and the experiments
were repeated three times to ensure reproducibility.

Seeger F., Quintyn R., Tanimoto A., Williams G.J., Tainer J.A., Wysocki V.H, & Garcin E.D. (2014). Interfacial Residues Promote an Optimal Alignment of the Catalytic Center in Human Soluble Guanylate Cyclase: Heterodimerization Is Required but Not Sufficient for Activity. Biochemistry, 53(13), 2153-2165.

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