Under nitrogen atmosphere, medium was replaced by PBS containing 1 mM of the phosphodiesterase inhibitor isobutylmethylxanthin (IBMX) to prevent cGMP degradation. After nitrite addition and 1 hour incubation the supernatant was removed and the adherent cells lysed by sonification in 500 µL lysis reagent provided with the enzyme immunoassay for detection of cGMP (cGMP EIA, GE Healthcare, Fairfield, USA). Protein content of the lysates was determined by BCA protein assay (Pierce, Rockford, USA). The cell lysate was used to analyse cGMP. cGMP levels were measured in acetylated samples according to protocol 3 of the manufacturer’s instructions. Optical density (405 nm) was measured by a plate reader (Tecan, Männedorf, Switzerland) and the concentration of cGMP was calculated from a standard curve produced from serial dilutions of acetylated cGMP solutions. All standards and samples were measured in duplicates.
Cgmp eia
Manufactured by GE Healthcare
Sourced in United States
The CGMP EIA is a laboratory equipment product designed for the quantitative determination of analytes in various sample types. It utilizes an enzyme-linked immunosorbent assay (EIA) technique to detect and measure specific target molecules. The CGMP EIA provides a standardized and validated method for conducting these analyses in a controlled, Good Manufacturing Practice (GMP) environment.
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Lab products found in correlation
2 protocols using cgmp eia
1
Quantification of Cellular cGMP Levels
2
Quantifying Plasma and Urinary Nitric Oxide
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Insulin ELISAs were purchased from Mercodia (Uppsala, Sweden). Multiplex proinflammatory 7 cytokine kit and custom made metabolic kit (Glucagon, active GLP-1, Insulin, and Leptin) were purchased from Mesoscale Discoveries (Rockville, MD, USA). T3 and T4 ELISA kits were purchased from Calbiotech (Spring Valley, CA, USA). cGMP EIA was purchased from GE Healthcare. All kits were run according to manufacturers' instructions. Plasma nitrite and nitrate and urinary nitrate were analyzed by HPLC (ENO-20) and autosampler (840, EiCom, Kyoto, Japan). Plasma was extracted using methanol (1:2) then centrifuged for 10 min 4 °C 10g. Urine samples were initially diluted (1:50) using carrier solution containing 10% methanol. Nitrate and nitrite were separated by reverse phase/ion exchange chromatography followed by nitrate reduction to nitrite by cadmium and reduced copper. The nitrite was then derivatized using Griess reagent to form diazo compounds and analyzed by detection at 540 nm.
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