1、RepSoxCat. No.: HY-13012CAS No.: 446859-33-2Molecular Formula: CHNMolecular Weight: 287.32Target: TGF- ReceptorPathway: TGF-beta/SmadStorage: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 monthSolvent NaCl, 100 mM; MgCl2, 5 mM; MnCl2, 5 mM; and DTT, 10 mM). The enzyme is preincubated w
2、ith different concentrations of RepSox (0.1% DMSO final concentration in the test) for 10 min at 37C. The reaction is then initiated by the addition of 3 M ATP (0.5 Ci -33P-ATP). After 15 min at 37C, phosphorylation is stopped by the addition of SDSPAGE sample buffer (50 mM Tris-HCl, pH 6.9, 2.5% gl
3、ycerol, 1% SDS, and 5% -mercaptoethanol). The samples are boiled for 5 min at 95C and run on a 12% SDSPAGE. Dried gels are exposed to a phosphor screen overnight. ALK5 autophosphorylation is quantified using a Storm imaging system1.MCE has not independently confirmed the accuracy of these methods. T
4、hey are for reference only.Cell Assay 1 To test anti-TGF- activity of compounds, HepG2 cells are seeded in 96 well microplates at a concentration of 35000 cells per well in 200 L of serum-containing medium. The microplates are then placed for 24 h in a cell incubator at 37C, 5% CO2 atm. RepSox disso
5、lved in DMSO are then added at concentrations of 50 nM to 10 M (final concentration of DMSO 1%) for 30 min prior to the addition of recombinant TGF- (1 ng/mL). After an overnight incubation, the cells are washed with PBS and lysed by addition of 10 L of passive lysis buffer. Inhibition of luciferase
6、 activity relative to control groups is used as a measure of compound activity. A concentrationresponse curve is constructed from which an IC50 value is determined graphically1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.CUSTOMER VALIDATIONSee more
7、customer validations on www.MedChemEREFERENCES1. Gellibert F, et al. Identification of 1,5-naphthyridine derivatives as a novel series of potent and selective TGF-beta type I receptor inhibitors. J Med Chem. 2004 Aug 26;47(18):4494-506.2. Ichida JK, et al. A small-molecule inhibitor of tgf-Beta sign
8、aling replaces sox2 in reprogramming by inducing nanog. Cell Stem Cell. 2009 Nov 6;5(5):491-503.Caution: Product has not been fully validated for medical applications. For research use only.Tel: 609-228-6898 Fax: 609-228-5909 E-mail: techMedChemEAddress: 1 Deer Park Dr, Suite Q, Monmouth Junction, NJ 08852, USA Oncotarget. 2017 Jun 19;8(36):60159-60172.www.MedChemE2
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