Western blot analysis of HT-29 cells, untreated (-) or treated with combinations of the following treatments as indicated: Z-VAD(OMe)-FMK #60332 (Z-VAD, 20 μM, added 30 min prior to other compounds; +), human TNF-α #8902 (hTNF-α, 20 ng/ml, 7 hr; +), SM-164 (100 nM, 7 hr; +), and necrostatin-1s (Nec-1s, indicated concentrations, 7 hr), using Phospho-RIP (Ser166) (D1L3S) Rabbit mAb #65746 (upper), RIP (D94C12) XP® Rabbit mAb #3493 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Chemical structure of Necrostatin-1s.
Necrostatin-1s is supplied as a lyophilized powder. For a 20 mM stock, reconstitute 5 mg of powder in 900 μl of DMSO. Working concentrations and length of treatment can vary depending on the desired effect.
Store lyophilized at room temperature, desiccated. In lyophilized form, the chemical is stable for 24 months. Once in solution, store at -20ºC and use within 3 months to prevent loss of potency. Aliquot to avoid multiple freeze/thaw cycles.
Molecular Weight: 277.7 g/mol
Molecular Formula: C13H12CIN3O2
Solubility: Soluble in DMSO at 25 mg/ml.
Necrostatin-1s, also known as Nec-1s or 7-Cl-O-Nec-1, is a potent and selective inhibitor of necroptosis. This analog of Necrostatin-1 (Nec-1) has been shown to be a specific receptor-interacting protein kinase (RIPK) 1 inhibitor that is metabolically more stable than the other variants (1). Research studies have found that Nec-1s is more selective than Nec-1 and Nec-1i (inactive variant) due to the fact that both molecules have an inhibitory affect on indoleamine 2,3-dioxygenase (IDO), a potent immunosuppressive molecule (2). Evidence also suggests that Nec-1s is around two-times more effective at RIPK1 inhibition than Nec-1 (IC50 = 210 nM vs. 494 nM) (1).
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