| 名稱 | JNK-IN-8 |
| 描述 | JNK-IN-8 (JNK Inhibitor XVI) is an irreversible JNK1/2/4 inhibitor (IC50: 4.7/18.7/1 nM). The selectivity is higher 10-fold than MNK2, Fms and no inhibition of Met, c-Kit, PDGFRβ in A375 cell line. |
| 細(xì)胞實(shí)驗(yàn) | JNK-IN-8 is dissolved in DMSO and stored, and then diluted with appropriate media before use[1]. HEK-293 cells stably expressing Interleukin Receptor 1 (HEK293-IL1R) are cultured in Dulbecco's Modified Eagle's medium (DMEM) supplemented with 10% FBS, 2 mM glutamine and 1×antimycotic/antibiotic solution. Cells are serum starved for 18 h before incubation with DMSO or JNK-IN-8, stimulated with 2 μM Anisomycin for 1h and lysates are clarified by centrifugation for 10 min at 16000 g and 4°C[1]. |
| 激酶實(shí)驗(yàn) | A375 cells are pre-treated with 1 μM JNK-IN-8 for the indicated amounts of time. Remove the medium and wash 3 times with PBS. Resuspend the cell pellet with 1 mL Lysis Buffer (1% NP-40, 1% CHAPS, 25 mM Tris, 150 mM NaCl, Phosphatase Inhibitor Cocktail, and Protease Inhibitor Cocktail). Rotate end-to-end for 30 min at 4°C. Lysates are cleared by centrifugation at 14000 rpm for 15 min in the Eppendorf. The cleared lysates gel filtered into Kinase Buffer (0.1% NP-40, 20 mM HEPES, 150 mM NaCl, Phosphatase Inhibitor Cocktail, Protease Inhibitor Cocktail) using Bio-Rad 10DG colums. The total protein concentration of the gel-filtered lysate should be around 5-15 mg/mL. Cell lysate is labeled with the probe from ActivX at 5 μM for 1 hour. Samples are reduced with DTT, and cysteines are blocked with iodoacetamide and gel filtered to remove excess reagents and exchange the buffer. Add 1 volume of 2X Binding Buffer (2% Triton-100, 1% NP-40, 2 mM EDTA, 2X PBS) and 50 μL streptavidin bead slurry and rotate end-to-end for 2 hours, centrifuge at 7000 rpm for 2 min. Wash 3 times with 1X Binding Buffer and 3 times with PBS. Add 30 μL 1X sample buffer to beads, heat samples at 95°C for 10 min. Run samples on an SDS-PAGE gel at 110V. After transferred, the membrane is immunoblotted with JNK antibody[1]. |
| 體內(nèi)活性 | JNK-IN-8（10 mM）對(duì)IL-1R細(xì)胞中IL-1β刺激的c-Jun磷酸化有抑制作用。與伊馬替尼相比,JNK-IN-8有明顯的1,4-雙苯胺和1,3-氨基苯甲酸結(jié)構(gòu)區(qū)域選擇性,且以N,N-二甲基丁烯乙酰胺共價(jià)結(jié)合Cys154靶點(diǎn)。JNK-IN-8抑制HeLa（EC50：486 nM）和A375（EC50：338 nM）細(xì)胞中c-Jun的磷酸化。JNK-IN-8與PIK3C3,IRAK1,PIP5K3和PIP4K2C結(jié)合可使選擇性和消除率顯著提高。JNK-IN-8經(jīng)Cys116抑制JNK2。 |
| 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 : 55 mg/mL (108.36 mM)
H2O : < 1 mg/mL (insoluble or slightly soluble)
Ethanol : < 1 mg/mL (insoluble or slightly soluble)
|
| 關(guān)鍵字 | JNKIN8 | Inhibitor | JNK | JNK IN 8 | inhibit | JNK-IN-8 |
| 相關(guān)產(chǎn)品 | Astragaloside IV | (-)-Bornyl acetate | Imatinib | Gilteritinib | Anisomycin | Axitinib | Sorafenib tosylate | Lenvatinib mesylate | Regorafenib | Pazopanib | Sorafenib | Regorafenib monohydrate |
| 相關(guān)庫 | 抑制劑庫 | 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 | 高選擇性抑制劑庫 | 神經(jīng)退行性疾病化合物庫 | 膜蛋白靶向化合物庫 | 酪氨酸激酶分子庫 | 疼痛相關(guān)化合物庫 |