| 名稱 | GSK 3 Inhibitor IX |
| 描述 | GSK 3 Inhibitor IX (6-BIO) is a selective reversible, ATP-competitive inhibitor of GSK-3α/β and CDK1-cyclinB complex. It inhibits (GSK-3α/β)/CDK1/CDK5 activity with IC50 values of 5 nM/320 nM/83 nM, respectively. |
| 細(xì)胞實(shí)驗(yàn) | COS1, Hepa (wild-type, CEM/LM AhR deficient and ELB1 ARNT deficient), or SH-SY5Y cells are grown in 6 cm culture dishes in Dulbecco's Modified Medium (DMEM) containing 10% fetal bovine serum. For treatment, IO (5 μM), BIO (5 or 10 μM), MeBIO (5 or 50 μM), LiCl (20 or 40 mM), or mock solution (DMSO, 0.5% final concentration) is added to medium when cell density reaches ～70% confluence. After 12 (SH-SY5Y) or 24 hours, the cells, while still in plate, are lysed with lysis buffer (1% SDS, 1 mM sodium orthovanadate, 10 mM Tris [pH 7.4]). The lysate is passed several times through a 26 g needle, centrifuged at 10,000 × g for 5 min, and adjusted to equal protein concentration. About 8 μg of each sample is loaded for immunoblotting. Enhanced chemiluminescence is used for detection. The following primary antibodies are used: mouse anti-β-catenin CT (Upstate Biotechnolgies, Clone 7D8, recognizes total β-catenin), mouse anti-phospho-β-catenin (Upstate Biotechnologies, Clone 8E7, recognizes dephosphorylated β-catenin), mouse anti-GSK-3 β, mouse anti-GSK-3 phosphoTyr216, rabbit anti-AhR (Aryl hydrocarbon receptor), and rabbit anti-actin.(Only for Reference) |
| 激酶實(shí)驗(yàn) | Kinase assay: Kinase activities are assayed in Buffer A or C at 30°C, at a final ATP concentration of 15 μM. Blank values are subtracted and activities calculated as pmoles of phosphate incorporated during a 10 min incubation. Controls are performed with appropriate dilutions of dimethylsulfoxide. In a few cases phosphorylation of the substrate is assessed by autoradiography after SDS-PAGE. GSK-3α/β is purified from porcine brain by affinity chromatography on immobilized axin. It is assayed, following a 1/100 dilution in 1 mg BSA/ml 10 mM DTT, with 5 μl 40 μM GS-1 peptide, a specific GSK-3 substrate, (YRRAAVPPSPSLSRHSSPHQSpEDEEE), in buffer A, in the presence of 15 μM [γ-32P] ATP (3,000 Ci/mmol; 1 mCi/ml) in a final volume of 30 μl. After 30 min incubation at 30°C, 25 μl aliquots of supernatant are spotted onto 2.5 × 3 cm pieces of Whatman P81 phosphocellulose paper, and 20 seconds later, the filters are washed five times (for at least 5 min each time) in a solution of 10 ml phosphoric acid/liter of water. The wet filters are counted in the presence of 1 ml ACS scintillation fluid. |
| 體外活性 | BIO抑制小鼠移植瘤模型中黑色素瘤生長. |
| 體內(nèi)活性 | BIO是pan-JAK抑制劑,作用于TYK2,JAK1,JAK2 和 JAK3,IC50值分別為0.03,1.5,8.0,0.5 μM。BIO是糖原合成酶激酶-3（GSK-3）的特異性抑制劑,GSK-3α/β的IC50為5 nM,比CDK5選擇性高16倍以上。BIO與這些激酶的ATP結(jié)合口袋相互作用,減少細(xì)胞模型中GSK-3特異性位點(diǎn)上的β-連環(huán)蛋白磷酸化。BIO促進(jìn)哺乳動(dòng)物心肌細(xì)胞增殖。BIO選擇性抑制STAT3磷酸化,并誘導(dǎo)人類黑色素瘤細(xì)胞凋亡。在人類和小鼠胚胎干細(xì)胞中,BIO維持未分化表型并維持多能狀態(tài)特異性轉(zhuǎn)錄因子Oct-3/4,Rex-1和Nanog的表達(dá)。 |
| 存儲(chǔ)條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | DMSO : 45 mg/mL (126.34 mM)
Ethanol : 21 mg/mL (58.96 mM)
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| 關(guān)鍵字 | Cyclin dependent kinase | Apoptosis | Inhibitor | Glycogen synthase kinase-3 | CDK | Glycogen synthase kinase 3 | MLS-2052 | MLS2052 | GSK-3 | inhibit | GSK 3 Inhibitor IX |
| 相關(guān)產(chǎn)品 | L-Glutamic acid | Gefitinib | Metronidazole | 5-Fluorouracil | Dextran sulfate sodium salt (MW 4500-5500) | Stavudine | Tributyrin | Myricetin | L-Ascorbic acid | Acetylcysteine | Salicylic acid | Sodium 4-phenylbutyrate |
| 相關(guān)庫 | 抑制劑庫 | 神經(jīng)保護(hù)化合物庫 | 經(jīng)典已知活性庫 | 已知活性化合物庫 | 激酶抑制劑庫 | 抗衰老化合物庫 | 酪氨酸激酶分子庫 | 藥物功能重定位化合物庫 | 抗癌臨床化合物庫 | 抗癌藥物庫 |