RAW264.7 cells were cultured for 4 h in black 96-well plates at a density of 1 × 105 cells/well and then treated with β-glucan or Euglena for 48 h. NO production was measured using a Nitric Oxide Synthase Kit obtained from Sigma‒Aldrich (USA) according to the manufacturer’s protocol. The fluorescence intensity was measured using a SpectraMax i3x plate reader (Molecular Devices, USA) with an excitation wavelength of 490 nm and an emission wavelength of 520 nm.
Nitric oxide synthase kit
Manufactured by Merck Group
Sourced in United States
The Nitric Oxide Synthase Kit is a laboratory tool designed to measure the activity of nitric oxide synthase (NOS) enzymes. NOS enzymes are responsible for the production of nitric oxide, a important signaling molecule in various biological processes. The kit provides the necessary reagents and protocols to quantify NOS activity through colorimetric or fluorometric detection methods.
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Lab products found in correlation
2 protocols using nitric oxide synthase kit
1
Quantification of Nitric Oxide in RAW264.7 Cells
2
Intracellular Nitric Oxide Measurement
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Intracellular nitric oxide was measured in living cells under physiological conditions by nitric oxide synthase kit (Sigma, St. Louis, MO, USA). The detection system utilizes a cellpermeable diacetate derivative of 4,5-diaminofluorescein (DAF-2 DA). DAF-2 DA penetrates cells rapidly, where it is hydrolyzed by intracellular esterase activity to DAF-2 that, in turn, reacts with NO produced by nitric synthase to form a fluorescent triazolofluorescein. The resulted fluorescence (excitation: 490 nm and emission: 520 nm) was detected by the plate reader (Kojima et al. 1998; (link)Navarro-Antolin and Lamas 2001) (link). The cells were seeded in black bottom 96-well plates at a density of 50,000 cell/well and allowed to attach for 24 h. After treatment with the IC 50 of the tested materials for 48 h, nitric oxide was detected according to the manufacture's procedure.
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