| 名稱 | ML-7 hydrochloride |
| 描述 | ML-7 hydrochloride (ML-7 HCl) is a cell-permeable, reversible, effective, ATP-competitive, and specific inhibitor of myosin light chain kinase (Ki: 300 nM); also inhibits smooth-muscle myosin light chain kinase, PKA, and PKC. |
| 細(xì)胞實(shí)驗(yàn) | Cells are treated with vehicle alone, Blebbistatin, ML-7, ML-9, latrunculin A or Y-27632 in media without serum at varying concentrations and time points before being fixed or prepared for FACS analysis. Untreated cells are also serum starved for the duration of the inhibitor treatment. Stock solutions of each agent are made in DMSO (50 μM Blebbistatin, 5 mM LA), 50% ethanol (10 mM ML-7), 70% ethanol (20 mM ML-9) or water (20 mM Y-27632) and are maintained at -20°C.(Only for Reference) |
| 激酶實(shí)驗(yàn) | IKK-2 Assay: Recombinant human IKK-2 (residues 1-756) is expressed in baculovirus as an N-terminal GST-tagged fusion protein, and its activity is assessed using a time-resolved fluorescence resonance energy transfer assay. In brief, IKK-2 (5 nM final) diluted in assay buffer (50 mM HEPES, 10 mM MgCl2, 1 mM CHAPS, pH 7.4, with 1 mM DTT and 0.01% w/v BSA) is added to wells containing various concentrations of compound or dimethyl sulfoxide (DMSO) vehicle (3% final). The reaction is initiated by the addition of GST-IκBα substrate (25 nM final)/ATP (1 μM final), in a total volume of 30 μL. The reaction is incubated for 30 min at room temperature, then terminated by the addition of 15 μL of 50 mM EDTA. Detection reagent (15 μL) in buffer (100 mM HEPES, pH 7.4, 150 mM NaCl, and 0.1% w/v BSA) containing antiphosphoserine- IκBα-32/36 monoclonal antibody 12C2, labeled with W-1024 europium chelate, and an allophycocyanin-labeled anti-GST antibody is added, and the reaction is further incubated for 60 min at room temperature. The degree of phosphorylation of GST- IκBαis measured as a ratio of specific 665-nm energy transfer signal to reference europium 620-nm signal, using a Packard Discovery plate reader. |
| 體外活性 | ML-7通過抑制MLCK激活粘附性MCF-10A及其Ras變異細(xì)胞中的caspase-3[2]。ML-7處理以劑量依賴的方式降低MLC20磷酸化水平，并相應(yīng)增加平滑肌細(xì)胞(SMC)的死亡率。ML-7對(duì)MLCK的抑制作用具有高度選擇性，其對(duì)MLCK的Ki為0.3 μM，而對(duì)蛋白激酶A的Ki為21 μM，對(duì)蛋白激酶C的Ki為42 μM。過表達(dá)Bcl-2可以保護(hù)細(xì)胞免受ML-7引起的凋亡[4]。 |
| 體內(nèi)活性 | ML7通過調(diào)節(jié)高脂飲食喂養(yǎng)的兔子中的MLCK和MLC磷酸化作用，影響緊密連接（TJ）蛋白區(qū)域橋蛋白（ZO)-1和閉鎖蛋白的表達(dá)，從而改善血管內(nèi)皮功能障礙（VED）和動(dòng)脈粥樣硬化（AS）。ML7能降低高脂飲食兔子動(dòng)脈壁中MLCK和MLC的磷酸化表達(dá)，并減少AS兔子中的脂質(zhì)沉積損傷[1]。 |
| 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 : 60 mg/mL (132.53 mM), Sonication is recommended.
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| 關(guān)鍵字 | ML-7 Hydrochloride | Inhibitor | Yes-associated protein | inhibit | Myosin | ML-7 | ML7 hydrochloride | ML7 | ML7 Hydrochloride | YAP | ML 7 | ML 7 hydrochloride | ML 7 Hydrochloride |
| 相關(guān)產(chǎn)品 | JAK1/2/3 Inhibitor 1 | Camostat mesylate | AICAR | FOY 251 | Acefylline | Benzamidine hydrochloride | Staurosporine | α-Vitamin E | Nafamostat mesylate |
| 相關(guān)庫(kù) | 抑制劑庫(kù) | 經(jīng)典已知活性庫(kù) | 抗癌化合物庫(kù) | 已知活性化合物庫(kù) | 細(xì)胞周期化合物庫(kù) | 激酶抑制劑庫(kù) | 抗衰老化合物庫(kù) | 干細(xì)胞分化化合物庫(kù) | TGF-β/Smad靶點(diǎn)化合物庫(kù) | 酪氨酸激酶分子庫(kù) |