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Cyclic gmp elisa kit

Manufactured by Cayman Chemical
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The Cyclic GMP ELISA Kit is a quantitative enzyme-linked immunosorbent assay (ELISA) designed for the measurement of cyclic guanosine monophosphate (cGMP) levels in biological samples. The kit utilizes a competitive ELISA format and includes all the necessary reagents and materials for the assay.

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Lab products found in correlation

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ELISAs (3 citations) Cyclic di-GMP ELISA Kit (3 citations) Cyclic GMP kit (2 citations) GMP ELISA Kit (2 citations) Cyclic GMP competitive ELISA Kit (2 citations)

14 protocols using cyclic gmp elisa kit

1

Quantifying cGMP Levels in Retinal Organoids

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cGMP concentrations were calculated for individual whole ROs by ELISA (Cyclic GMP ELISA kit, Cayman Chemicals, Ann Arbor, MI, USA) according to the manufacturer’s instructions. In brief, ROs were washed with PBS then incubated in 100 μL 0.1M HCl for 20 min at RT and mechanically homogenized. The samples were centrifuged at 1,000×g for 10 min then the supernatant combined with 200 μL ELISA buffer. We used 50 μL of this sample per well. Absorbance was measured at a wavelength of 420 nm. Values given are normalized to volume to give cGMP concentration per whole RO with at least three RO analysed per condition, and statistical significance calculated by one-way ANOVA with Tukey’s HSD post hoc analysis where ∗, ∗∗, and ∗∗∗ denote a p value ≤ 0.05, ≤0.01, and ≤0.005, respectively.
Sai H., Ollington B., Rezek F.O., Chai N., Lane A., Georgiadis T., Bainbridge J., Michaelides M., Sacristan-Reviriego A., Perdigão P.R., Leung A, & van der Spuy J. (2024). Effective AAV-mediated gene replacement therapy in retinal organoids modeling AIPL1-associated LCA4. Molecular Therapy. Nucleic Acids, 35(1), 102148.
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2

Cyclic GMP Quantification in Plasma and Erythrocytes

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To assess plasmatic and intra-erythrocytic cGMP levels, in a subgroup of 10 subjects of each condition (CTRL, CT, PD, HD) plasma and RCBs were processed to obtain suitable samples for the enzyme-linked immunosorbent assay (ELISA). Regarding the plasmatic fraction, it was treated with absolute ethanol whereas RBCs were washed thrice with a washing solution (Glucose 10 mM and BSA 10 mg/mL in PBS w/o Ca++/Mg++) to remove potential mononuclear cells residues and HCl 1N was then added. Both plasma and erythrocytes samples were successively dry-frozen. Then, 500 µL of plasma and 500 µL of RBCs were processed following the manufacturers’ instructions (Cyclic GMP ELISA Kit, Cayman Chemical Company, Ann Arbor, MI, USA).
The absorbance at 520 nm was read by using a microplate reader (SpectraMAX 190, Molecular Devices, Sunnyvale, CA, USA) and it was inversely proportional to the amount of cGMP contained in the analyzed samples. cGMP concentration was calculated using a standard curve as reference and data were expressed as pmol/mL.
Palmerini C., Piscitani L., Bologna G., Riganti C., Lanuti P., Mandatori D., Di Liberato L., Di Fulvio G., Sirolli V., Renda G., Pipino C., Marchisio M., Bonomini M., Pandolfi A, & Di Pietro N. (2021). Predialysis and Dialysis Therapies Differently Affect Nitric Oxide Synthetic Pathway in Red Blood Cells from Uremic Patients: Focus on Peritoneal Dialysis. International Journal of Molecular Sciences, 22(6), 3049.
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3

Quantifying cGMP Levels via ELISA

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The cGMP was measured using the Cyclic GMP ELISA Kit (Cayman Chemical, Ann Arbor, MI, USA) following the manufacturer's instructions.
Nishiguchi Y., Hata Y., Date R., Fujimoto D., Umemoto S., Kanki T., Yokoi H., Mori K.P., Handa T., Watanabe-Takano H., Kanai Y., Yasoda A., Izumi Y., Kakizoe Y., Mochizuki N., Mukoyama M, & Kuwabara T. (2021). Osteocrin, a bone-derived humoral factor, exerts a renoprotective role in ischemia–reperfusion injury in mice. Nephrology Dialysis Transplantation, 37(3), 444-453.
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4

Measurement of Urinary cGMP Levels

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The concentration of cGMP in the urine was measured using the Cayman Cyclic GMP ELISA kit. Urine samples were diluted 1:7,500 in ELISA buffer and assayed directly following the manufacturer’s instructions. Values of the samples were interpolated from the standard curve using GraphPad Prism software.
Golshiri K., Ataabadi E.A., Jüttner A.A., Snyder G.L., Davis R.E., Lin A., Zhang L., de Vries R., Garrelds I.M., Leijten F.P., Danser A.H, & Roks A.J. (2022). The Effects of Acute and Chronic Selective Phosphodiesterase 1 Inhibition on Smooth Muscle Cell-Associated Aging Features. Frontiers in Pharmacology, 12, 818355.
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5

Measuring CNP and OSTN-Induced cGMP in Ocy454 Cells

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Ocy454 cells, grown in 96-well plates, were incubated at 37 °C for 2 days. Cells were subsequently stimulated with an incubation medium (Hank’s Balanced Salt solution with 0.075% BSA, 10 mM HEPES, and 2 mM IBMX) containing 50 nM human CNP or 500 nM human OSTN at room temperature for 30 min. Cells were collected in 0.1 M HCl. The amount of cGMP in each cell lysate was measured using the Cyclic GMP ELISA Kit (Cayman Chemical) according to the manufacturer’s protocol.
Wang J.S., Kamath T., Mazur C.M., Mirzamohammadi F., Rotter D., Hojo H., Castro C.D., Tokavanich N., Patel R., Govea N., Enishi T., Wu Y., da Silva Martins J., Bruce M., Brooks D.J., Bouxsein M.L., Tokarz D., Lin C.P., Abdul A., Macosko E.Z., Fiscaletti M., Munns C.F., Ryder P., Kost-Alimova M., Byrne P., Cimini B., Fujiwara M., Kronenberg H.M, & Wein M.N. (2021). Control of osteocyte dendrite formation by Sp7 and its target gene osteocrin. Nature Communications, 12, 6271.
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6

Quantifying Plasma Nitric Oxide and cGMP Levels

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Blood was collected in EDTA tubes and spun at 3000 x g for 10 minutes at 4o C. The collected plasma was assessed in duplicate using the Nitrate/Nitrite Colorimetric Assay Kit (Catalog# 78001, Cayman Chemical, Ann Arbor, MI) according to the manufacturer’s instructions. Levels of cGMP were measured in plasma and placental homogenates in duplicate using the Cyclic GMP ELISA Kit (# 581021, Cayman Chemical) according to the manufacturer’s instructions. The protein concentration of the placental homogenates was measured using a protein assay with BSA standards. All placental data were normalized to protein and are expressed as pmol/mg.
Travis O.K., Baik C., Tardo G.A., Amaral L., Jackson C., Greer M., Giachelli C., Ibrahim T., Herrock O.T., Williams J.M, & Cornelius D.C. (2020). Adoptive Transfer of Placental Ischemia Stimulated Natural Killer Cells Causes a Preeclampsia-Like Phenotype in Pregnant Rats. American journal of reproductive immunology (New York, N.Y. : 1989).
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7

cGMP Quantification by ELISA

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We used the Cyclic GMP ELISA Kit (Cayman Chemical, cat#581021) according to manufacturer’s instructions and final cGMP quantification was normalized to total protein.
Maretti-Mira A.C., Wang X., Wang L, & DeLeve L.D. (2019). Incomplete differentiation of engrafted bone marrow endothelial progenitor cells initiates hepatic fibrosis in the rat. Hepatology (Baltimore, Md.), 69(3), 1259-1272.
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8

Evaluating CM414 Treatment in Mdr2-KO Mice

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Female Mdr2-KO− and Mdr2-WT mice (The Jackson Laboratory, BarHarbor, ME, USA) fed a standard laboratory diet were used in this study. Two littermates groups of 6-month-old Mdr2-KO mice were established. One group was treated daily with CM414 (40 mg/kg, i.p.) (n = 5) or vehicle (10% DMSO, 10% Tween-20 in saline solution) (n = 5) for 4 weeks. The control group of Mdr2-WT mice (n = 8) received the same volume of vehicle. This dose of CM414 was selected from a previous study [56 (link)]. At the end of treatments serum was withdrawn and animals were euthanized. Livers were excised, weighted, and were either span frozen of fixed in formalin and paraffin-embedded. Serums levels of liver enzymes levels were analyzed in a Cobas analyzer (Roche, Mannheim, Germany) as previously described [20 (link),116 (link),117 (link),118 (link)]. cGMP levels in mouse liver tissues were measured with the Cyclic GMP ELISA kit from Cayman Chemical (581021) as recommended by the supplier. The study was approved by the Ethics Committee for Animal Experimentation of the University of Navarra (Protocol number 086-19).
Claveria-Cabello A., Colyn L., Uriarte I., Latasa M.U., Arechederra M., Herranz J.M., Alvarez L., Urman J.M., Martinez-Chantar M.L., Banales J.M., Sangro B., Rombouts K., Oyarzabal J., Marin J.J., Berasain C., Avila M.A, & Fernandez-Barrena M.G. (2020). Dual Pharmacological Targeting of HDACs and PDE5 Inhibits Liver Disease Progression in a Mouse Model of Biliary Inflammation and Fibrosis. Cancers, 12(12), 3748.
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9

Placental Superoxide Production Assay

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Superoxide production in the placenta was measured using the lucigenin technique, as previously described by our lab.[23 (link), 28 , 29 (link)] Briefly, placentas chosen at random from all groups were snap frozen in liquid nitrogen immediately after collection and stored at −80 °C until further processing. Placentas were homogenized using the Bio-Rad Cell Lysis Kit (Bio-Rad, Hercules, CA) according to the manufacturer’s instructions. The tissue lysate was incubated with lucigenin (Sigma-Aldrich, St. Louis, MO) at a concentration of 5 μM. The samples were allowed to equilibrate for 15 min in the dark, and the luminescence was measured for 10 s with a BioTek Plate Reader (BioTek, Winooski, VT). Luminescence was recorded as relative light units per minute (RLUs/min). An assay blank containing lucigenin with no homogenate was subtracted from the reading before transformation of the data. Each sample was run in triplicate and the average was used for data transformation. Levels of cGMP were measured in placental homogenates in duplicate using the Cyclic GMP ELISA Kit (# 581021, Cayman Chemical) according to the manufacturer’s instructions. The protein concentration of placental homogenates was measured using a protein assay with BSA standards (Pierce, Rockford, IL). All placental data were normalized to protein concentration and are expressed as RLU/min/mg protein.
Travis O.K., Tardo G.A., Giachelli C., Siddiq S., Nguyen H.T., Tate Crosby M., Johnson T., Brown A.K., Williams J.M, & Cornelius D.C. (2021). Tumor Necrosis Factor-Alpha Blockade Improves Uterine Artery Resistance, Maternal Blood Pressure, and Fetal Growth in Placental Ischemic Rats. Pregnancy hypertension, 25, 39-47.
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10

Activation of soluble guanylate cyclase

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Pure human recombinant soluble guanylate cyclase was purchased from Enzo Lifesciences. Apo-sGC was prepared as previously described30 (link). NO-ferroheme-Mb (10 µM) or DEA-NONOate (10 µM) was added to apo-sGC (~1 µM) dissolved in medium containing the following: (1) 10 mM GTP, (2) 10 mM HEPES, (3) 150 mM NaCl, (4) 1 mM DTT and (5) 3 mM MgCl2. The Eppendorf tubes were placed in a heated water bath (37 °C) for 10 min. sGC was inactivated by moving the Eppendorf tubes to a heat block (100 °C) for 3 min. The samples were stored at −80 °C until analysis. cGMP was measured using a commercial ELISA kit (Cayman Chemical; cyclic GMP ELISA kit).
Kleschyov A.L., Zhuge Z., Schiffer T.A., Guimarães D.D., Zhang G., Montenegro M.F., Tesse A., Weitzberg E., Carlström M, & Lundberg J.O. (2023). NO-ferroheme is a signaling entity in the vasculature. Nature Chemical Biology, 19(10), 1267-1275.
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