| 名稱 | Tubastatin A |
| 描述 | Tubastatin A (Tubastatin A BASE) is an effective and specific HDAC6 inhibitor (IC50: 15 nM, in a cell-free assay). Its selectivity is 1000-fold against all the other isozymes except HDAC8. |
| 細(xì)胞實驗 | Anchorage-independent growth is assessed by growing cells in soft agar. About 25,000 cells suspended in 0.4% agar in culture media are layered over a 1% agar layer in a 6-well plate. Media are added twice a week and pictures are taken after 21 days of incubation. The number and size of colonies are analyzed using the Gel-Pro software.(Only for Reference) |
| 激酶實驗 | HDAC enzymatic assays: Tubastatin A is dissolved and diluted in assay buffer (50 mM HEPES, pH 7.4, 100 mM KCl, 0.001% Tween-20, 0.05% BSA, and 20 μM tris(2-carboxyethyl)phosphine) to 6-fold of the final concentration. HDAC enzymes are diluted to 1.5-fold of the final concentration in assay buffer and pre-incubated with Tubastatin A for 10 minutes before the addition of the substrate. The amount of FTS (HDAC1, HDAC2, HDAC3, and HDAC6) or MAZ-1675 (HDAC4, HDAC5, HDAC7, HDAC8, and HDAC9) used for each enzyme is equal to the Michaelis constant (Km), as determined by a titration curve. FTS or MAZ-1675 is diluted in assay buffer to 6-fold the final concentration with 0.3 μM sequencing grade trypsin. The substrate/trypsin mix is added to the enzyme/compound mix and the plate is shaken for 60 seconds and then placed into a SpectraMax M5 microtiter plate reader. The enzymatic reaction is monitored for release of 7-amino-4-methoxy-coumarin over 30 minutes, after deacetylation of the lysine side chain in the peptide substrate, and the linear rate of the reaction is calculated. |
| 體外活性 | 與對照組相比,Tubastatin A 可使PCNA陽性細(xì)胞的量顯著降低(34% vs 65%).在同基因大鼠原位膽管癌模型中,Tubastatin A（10 mg/kg）降低瘤重與肝重和體重（分別為5和5.6倍）的比值.在給弗氏完全佐劑誘導(dǎo)的動物炎癥模型中,Tubastat A（30 mg/kg,i.p.）對鼠爪體積有顯著抑制作用.Tubastat A（30 mg/kg,i.p.）處理膠原誘導(dǎo)型關(guān)節(jié)炎DBA1小鼠的爪子組織,使臨床評分（70％）和IL-6表達(dá)顯著降低. |
| 體內(nèi)活性 | Tubastatin A（5 μM）劑量依賴性地使神經(jīng)元細(xì)胞免于同型半胱氨酸誘導(dǎo)的死亡,10 μM時可完全避免。 Tubastatin A（10 μM）對組蛋白乙?；休p微誘導(dǎo)作用。作用于膽管癌細(xì)胞系時,Tubastatin A（10 μM）促使乙?；?tubulin水平增加和初級纖毛表達(dá)的恢復(fù),初級纖毛表達(dá)的恢復(fù)與Hedgehog、MAPK信號通路下調(diào)、細(xì)胞增殖率（平均50％）和浸潤（40％）減少相關(guān)。作用于脂多糖刺激的人類 THP-1巨噬細(xì)胞時,Tubastatin A明顯抑制TNF-α和IL-6（IC50：272/712 nM）。作用于小鼠Raw 264.7巨噬細(xì)胞時,Tubastatin A劑量依賴性抑制一氧化氮分泌（IC50：4.2 μM）。 |
| 存儲條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | DMSO : 35 mg/mL (104.35 mM), Sonication is recommended.
Ethanol : < 1 mg/mL (insoluble or slightly soluble)
H2O : < 1 mg/mL (insoluble or slightly soluble)
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| 關(guān)鍵字 | Autophagy | Inhibitor | Apoptosis | inhibit | HDAC | Histone deacetylases | Tubastatin A |
| 相關(guān)產(chǎn)品 | Guanidine hydrochloride | Naringin | Valproic Acid | L-Glutamic acid | Gefitinib | Hydroxychloroquine | Dextran sulfate sodium salt (MW 4500-5500) | Stavudine | Tributyrin | L-Ascorbic acid | Paeonol | Sodium 4-phenylbutyrate |
| 相關(guān)庫 | 抑制劑庫 | 經(jīng)典已知活性庫 | 已知活性化合物庫 | 自噬庫 | 細(xì)胞凋亡化合物庫 | 抗衰老化合物庫 | 高選擇性抑制劑庫 | HIF-1化合物庫 | NF-κB 通路分子庫 | 細(xì)胞重編程化合物庫 |