SB-415286 - GSK-3 抑制劑 - 生命科學(xué)試劑 - MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemESB 415286Cat. No.: HY-15438CAS No.: 264218-23-7分式: CHClNO分量: 359.72作靶點(diǎn): GSK-3作通路: PI3K/Akt/mTOR; Stem Cell/Wnt儲(chǔ)存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實(shí)驗(yàn) DMSO : 83.3 mg/mL (231.57 mM)* means s

2、oluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制備儲(chǔ)備液1 mM 2.7799 mL 13.8997 mL 27.7994 mL5 mM 0.5560 mL 2.7799 mL 5.5599 mL10 mM 0.2780 mL 1.3900 mL 2.7799 mL請(qǐng)根據(jù)產(chǎn)品在不同溶劑中的溶解度，選擇合適的溶劑配制儲(chǔ)備液，并請(qǐng)注意儲(chǔ)備液的保存式和期限。BIOLOGICAL ACTIVITY物活性 SB 415286種有效的 GSK-3 抑制劑，IC50 值為 77.5 nM，Ki 值為 30.75

3、 nM；SB 415286 對(duì) GSK-3和 GSK-3 的抑制效果相同。IC50 & Target hGSK-3 hGSK-377.5 nM (IC50) 77.5 nM (IC50)1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemE體外研究 SB 415286 (SB-415286) inhibits human GSK-3 with an IC50 of 77.5 nM, and a Ki of 30.75 nM. SB-415286stimulates glycogen synthesis in the Chang human liver

4、 cell line with EC50 of 2.9 M. SB-415286 stimulatesglycogen synthase activity in Chang human liver cells. SB-415286 induces transcription of a -catenin-LEF/TCF regulated reporter gene in HEK293 cells 1. SB 415286 (SB-415286, 5-44 M) attenuates B65 cellloss mediated by 1 mM H2O2. SB-415286 (5-44 M) c

5、auses a significant dose-dependent decrease in thefluorescence intensity of DCF, and attenuates B65 ROS production as mediated by 1 mM H2O2. SB-415286(5-44 M) also attenuates ROS production in CGN mediated by 1 mM H2O2 2. SB-415286 (50 M) inducesa substantial suppression of immunoprecipitated GSK3 a

6、ctivity by 97% 3.PROTOCOLKinase Assay 1 GSK-3 kinase activity is measured, in the presence or absence of SB-216763 or SB-415286, in a reactionmixture containing final concentrations of: 1 nM human GSK-3 or rabbit GSK3; 50 mM MOPS pH 7.0; 0.2mM EDTA; 10 mM Mg-acetate; 7.5 mM -mercaptoethanol; 5% (w/v

7、) glycerol; 0.01% (w/v) Tween-20; 10%(v/v) DMSO; 28 M GS-2 peptide substrate. The GS-2 peptide sequence corresponds to a region ofglycogen synthase that is phosphorylated by GSK-3. The assay is initiated by the addition of 0.34 Ci 33P-ATP (IC50 determinations) or 2.7 Ci 33P-ATP (Ki determinations).

8、The total ATP concentration is 10 M(IC50 determinations) or ranges from 0 to 45 M (Ki determinations). Following 30 min incubation at roomtemperature the assay is stopped by the addition of one third assay volume of 2.5% (v/v) H3PO4 containing21 mM ATP. Samples are spotted onto P30 phosphocellulose

9、mats and these are washed six times in 0.5%(v/v) H3PO4. The filter mats are sealed into sample bags containing Wallac betaplate scintillation fluid. 33Pincorporation into the substrate peptide is determined by counting the mats in a Wallac microbeta scintillationcounter 1.MCE has not independently c

10、onfirmed the accuracy of these methods. They are for reference only.Cell Assay 2 B65 cells are used after 24 h of in vitro culture. CGN are used after 7-8 days in vitro. Lithium and SB-415286are dissolved in culture media and DMSO, respectively, and added to the neuronal preparation at the preciseco

11、ncentrations, 1 h before addition H2O2 (50 M to 1 mM). To assess the loss in cell viability, we use theMTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium method. MTT is added to the cells at a finalconcentration of 250 M and incubated for 1 h, allowing the reduction in MTT to produce a dark b

12、lueformazan product. Media are then removed, and cells are dissolved in dimethylsulfoxide. Formazanproduction is measured by the absorbency change at 595 nm using a microplate reader. Viability results areexpressed as percentages. The absorbency measured from non-treated cells is taken to be 100% 2.

13、MCE has not independently confirmed the accuracy of these methods. They are for reference only.戶使本產(chǎn)品發(fā)表的科研獻(xiàn) Oncotarget. 2017 Jul 7;8(47):82174-82184.See more customer validations on HYPERLINK / www.MedChemEREFERENCES2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemE1. Coghlan MP, et al. Selective smal

14、l molecule inhibitors of glycogen synthase kinase-3 modulate glycogen metabolism and genetranscription. Chem Biol. 2000 Oct;7(10):793-803.2. Pizarro JG, et al. Neuroprotective effects of SB-415286 on hydrogen peroxide-induced cell death in B65 rat neuroblastoma cells andneurons. Int J Dev Neurosci. 2008 May-Jun;26(3-4):269-76.3. MacAulay K, et al. Use of lithium and SB-415286 to explore the role of glycogen synthase kinase-3 in the regulation of glucose transportand glycogen synthase. Eur J Biochem. 2003