SKF-86002_DataSheet_MedChemExpress

Inhibitors, Agonists, Screening Libraries
Data Sheet
BIOLOGICAL ACTIVITY:
SKF–86002 is a potent inhibitor of p38 MAP kinase wit IC50 of 0.5–1 uM; inhibits LPS–induced IL–1 and TNF–α production in human monocytes (IC50 = 1 μM).
IC50 value:
Target: p38 MAPK inhibitor
in vitro: SKF–86002 inhibited prostaglandin H2 (PGH2) synthase activity (IC50 120 microM) as well as prostanoid production by rat basophilic leukemia (RBL–1) cells (IC50 70 microM) and its sonicate (IC50 100 microM) and human monocytes (IC50 1 microM). In addition, SK&F 86002 inhibited the generation of dihydroxyeicosatetraenoic acid (diHETE) and 5–hydroxyeicosatetraenoic acid
(5–HETE) by a high speed supernatant fraction of RBL–1 cells (IC50 10 microM) [1]. differentiation of HL–60 cells toward the neutrophil phenotype resulted in a loss in c–Jun NH2–terminal kinase activation with concomitant acquisition of
formylmethionylleucylphenylalanine–stimulatable and stress–inducible p38 MAPK activity as well as apoptosis blockade by SKF–86002
[2]. SKF–86002 blocked superoxide anion production in response to FMLP and reduced adhesion and chemotaxis in response to PAF or FMLP [3].
PROTOCOL (Extracted from published papers and Only for reference)
Enzyme asssay [3]
In vitro inhibition of MAPks were performed by adding a range
of concentrations of SKF–86002 to immunoprecipitated p38 MAPk or p42/44 (ERK) MAPk followed immediately by the kinase assays described above.Neutrophils resuspended at 20 × 106 /ml were incubated in complete KRPD for 30 min and stimulated with either FMLP or PAF as indicated in the text. Cell pellets were lysed in RIPA buffer and the triton–soluble lysates, precleared with protein
A–Sepharose, were immunoprecipitated in the presence of anti–p42/ 44 (ERK) MAPk. Protein A–Sepharose beads were washed once in RIPA buffer, twice in PAN and then subjected to an in vitro kinase assay with the addition of a 50 ml reaction mix (containing 50 mM b–glycerophosphate [pH 7.2], 100 mM Na3VO4, 20 mM MgCl2, 200 mM ATP, 50 mg/ml IP20, 1 mM EGTA, 400 mM EGFR661–681peptide, and 2 mCi [g–32P]–ATP) for 30 min at 308C. The reaction was terminated with a 10–s centrifugation at 5,000 rpm and the bead supernatant was treated in two ways: (a) 25 ml supernatant was added to tubes containing 25 ml 25% (wt/vol) TCA solution and 45 ml from mixture was spotted on P81 filter discs. These were washed three times in 75 mM phosphoric acid and once in acetone before being dried and counted in a b–counter; (b) 25 ml of 23 Laemmli sample buffer containing 40 mM DTT was added to the tubes containing the protein A–Sepharose beads and the remainder of the supernatant. Samples were boiled for 5 min at 1008C, subjected to SDS–PAGE, transferred to nitrocellulose and then probed with anti–phosphotyrosine and then anti–p42/44 (ERK) MAP kinase antibodies.
Product Name:
SKF–86002Cat. No.:
HY-12511CAS No.:
72873-74-6Molecular Formula:
C 16H 12FN 3S Molecular Weight:
297.35Target:
p38 MAPK Pathway:
MAPK/ERK Pathway Solubility:
10 mM in DMSO
References:
[1]. Griswold DE, et al. SK&F 86002: a structurally novel anti–inflammatory agent that inhibits lipoxygenase– and cyclooxygenase–mediated metabolism of arachidonic acid. Biochem Pharmacol. 1987 Oct 15;36(20):3463–70.
[2]. Frasch SC, et al. p38 mitogen–activated protein kinase–dependent and –independent intracellular signal transduction pathways leading to apoptosis in human neutrophils. J Biol Chem. 1998 Apr 3;273(14):8389–97.
[3]. Nick JA, et al. Common and distinct intracellular signaling pathways in human neutrophils utilized by platelet activating factor and FMLP. J Clin Invest. 1997 Mar 1;99(5):975–86.
Caution: Product has not been fully validated for medical applications. For research use only.
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