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Ns 398

Manufactured by Enzo Life Sciences
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Sourced in United States, Switzerland

NS-398 is a potent and selective inhibitor of cyclooxygenase-2 (COX-2), an enzyme involved in the production of prostaglandins. It exhibits an IC50 value of approximately 1.7 μM for the inhibition of COX-2 activity.

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Lab products found in correlation

Tnf α, by Enzo Life Sciences (1 mentions) Pge2 elisa kit, by Enzo Life Sciences (1 mentions) Zileuton, by Merck Group (1 mentions) Mk886, by Enzo Life Sciences (1 mentions)

4 protocols using ns 398

1

PGE2 Quantification in fAT-MSC Conditioned Media

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The cell-conditioned medium was obtained from fAT-MSCs, TNF-α-stimulated fAT-MSC (24 h), and TNF-α- and NS-398-stimulated (5 µM; Enzo Life Sciences) fAT-MSC (48 h) incubated in six-well plates at a density of 5 × 104 cells/well. Following the manufacturer’s instructions, a prostaglandin E2 (PGE2) ELISA kit (Enzo Life Sciences) was used to determine the concentration of PGE2 present in the cell-conditioned medium.
Kim K., An J.H., Park S.M., Lim G., Seo K.W, & Youn H.Y. (2023). Amelioration of DSS-induced colitis in mice by TNF-α-stimulated mesenchymal stem cells derived from feline adipose tissue via COX-2/PGE2 activation. Journal of Veterinary Science, 24(4), e52.
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2

Epithelial and Neutrophil Drug Treatments for Migration

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Epithelial drug treatments were performed immediately prior to infection. Epithelial cells were washed three times, and incubated with zileuton (Sigma-Aldrich; St. Louis, MO), CDC (Enzo Life Sciences), MK886 (Enzo Life Sciences), or NS398 (Enzo Life Sciences) at the indicated concentration for 1h at 37°C in HBSS. After drug treatment, epithelial cells were washed three times then infected, as above. Untreated neutrophils were then added for migration. In experiments calling for drug-treatment of neutrophils, neutrophils (5×107/mL) were suspended in zileuton, CDC, MK886 or vehicle controls at the indicated concentrations for 1h at 37°C in HBSS−. Immediately prior to migration, neutrophils were resuspended in HBSS containing inhibitors. Neutrophil viability was confirmed by trypan blue exclusion.
Pazos M.A., Pirzai W., Yonker L.M., Morisseau C., Gronert K, & Hurley B.P. (2014). Distinct cellular sources of Hepoxilin A3 and Leukotriene B4 are used to coordinate bacterial-induced neutrophil transepithelial migration. Journal of immunology (Baltimore, Md. : 1950), 194(3), 1304-1315.
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3

Modulating PGE2 in Anti-Inflammatory Response

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To assess the role of PGE2 produced by fAT-MSCs in the anti-inflammatory response, fAT-MSCs were treated with PGE2 inhibitor NS-398 (5 μM; Enzo Life Sciences, USA) for 24 h and then stimulated with INF-γ (50 ng/mL for 48 h). After that, the supernatant was collected and stored as described above.
Park S.G., An J.H., Li Q., Chae H.K., Park S.M., Lee J.H., Ahn J.O., Song W.J, & Youn H.Y. (2021). Feline adipose tissue-derived mesenchymal stem cells pretreated with IFN-γ enhance immunomodulatory effects through the PGE2 pathway. Journal of Veterinary Science, 22(2), e16.
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4

Measuring Nitric Oxide Release in Adipocytes

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Adipocytes kept in normoxia or hypoxia incubators were treated with or without TNFα (10 ng/ml) for 24 h. The cells were pre-treated either with iNOS specific inhibitor 1400W (10 µM; Cayman chemical) or COX-2 specific inhibitor NS398 (10µM; Enzo Life Sciences, Lausen, Switzerland). The culture supernatants were collected to measure the released nitric oxide (NO). Levels of NO, in form of nitrite (NO 2 -) were determined using Griess reagent (1% sulfanilamide, 0.1% N-1-naphthylenediamine dihydrochloride, and 2.5% phosphoric acid) as done previously [21] . The absorbance was measured at 570 nm with a multi-mode microplate reader (Molecular Devices, Spectra Max M2, Bucher Biotec, Basel, Switzerland).
Bhattacharya I., Domínguez A.P., Drägert K., Humar R., Haas E, & Battegay E.J. (2015). Hypoxia potentiates tumor necrosis factor-α induced expression of inducible nitric oxide synthase and cyclooxygenase-2 in white and brown adipocytes. Biochemical and biophysical research communications, 461(2).
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