| 名稱 | Tubastatin A Hydrochloride |
| 描述 | Tubastatin A Hydrochloride (TSA HCl) is an effective and specific HDAC6 inhibitor (IC50: 15 nM). It has selectivity (>1000-fold) against all other isozymes except HDAC8 (>57-fold). |
| 細(xì)胞實(shí)驗(yàn) | Primary cortical neuron cultures are obtained from the cerebral cortex of fetal Sprague-Dawley rats (embryonic day 17) as described previously. All experiments are initiated 24 hours after plating. Under these conditions, the cells are not susceptible to glutamate-mediated excitotoxicity. For cytotoxicity studies, cells are rinsed with warm PBS and then placed in minimum essential medium (Invitrogen) containing 5.5 g/L glucose, 10% fetal calf serum, 2 mM L-glutamine, and 100 μM cystine. Oxidative stress is induced by the addition of the glutamate analogue homocysteate (HCA; 5 mM) to the media. HCA is diluted from 100-fold concentrated solutions that are adjusted to pH 7.5. In combination with HCA, neurons are treated with Tubastatin A at the indicated concentrations. Viability is assessed after 24 hours by MTT assay (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) method.(Only for Reference) |
| 激酶實(shí)驗(yàn) | Enzyme Inhibition Assays: Enzyme inhibition assays are performed by the Reaction Biology Corporation, Malvern, PA, using the Reaction Biology HDAC Spectrum platform. (www.reactionbiology.com) The HDAC1, 2, 4, 5, 6, 7, 8, 9, 10, and 11 assays use isolated recombinant human protein; HDAC3/NcoR2 complex is used for the HDAC3 assay. Substrate for HDAC1, 2, 3, 6, 10, and 11 assays is a fluorogenic peptide from p53 residues 379-382 (RHKKAc); substrate for HDAC8 is fluorogenic diacyl peptide based on residues 379-382 of p53 (RHKAcKAc). Acetyl-Lys (trifluoroacetyl)-AMC substrate is used for HDAC4, 5, 7, and 9 assays. Tubastatin A is dissolved in DMSO and tested in 10-dose IC50 mode with 3-fold serial dilution starting at 30 μM. Control Compound Trichostatin A (TSA) is tested in a 10-dose IC50 with 3-fold serial dilution starting at 5 μM. IC50 values are extracted by curve-fitting the dose/response slopes. |
| 體外活性 | Tubastatin A 對(duì)所有11種HDAC異構(gòu)體具有較高選擇性，除HDAC8外對(duì)其他異構(gòu)體的選擇性超過1000倍，在HDAC8上大約有57倍的選擇性。在同型半胱氨酸(HCA)誘導(dǎo)的神經(jīng)退行性測(cè)定中，Tubastatin A 以劑量依賴的方式保護(hù)神經(jīng)細(xì)胞免受HCA誘導(dǎo)的細(xì)胞死亡，從5μM開始顯示保護(hù)效果，在10μM時(shí)幾乎完全保護(hù)。[1] 在體外實(shí)驗(yàn)中，Tubastatin A 在100 ng/mL的劑量下增加Foxp3+ 調(diào)節(jié)性T細(xì)胞（Tregs）對(duì)T細(xì)胞增殖的抑制。[2] 在C2C12細(xì)胞中，Tubastatin A 的處理會(huì)導(dǎo)致在肌肉生成過程早期時(shí)，由于α-微管蛋白過度乙酰化而妨礙肌管形成；然而，當(dāng)在肌管中α-微管蛋白過度乙酰化時(shí)，則會(huì)發(fā)生肌管伸長(zhǎng)。[3] 最近的研究表明，Tubastatin A 處理通過原子力顯微鏡（AFM）測(cè)試表明增加了細(xì)胞彈性，而不對(duì)小鼠卵巢癌細(xì)胞系MOSE-E和MOSE-L中的微絲或微管網(wǎng)絡(luò)造成嚴(yán)重變化。[4] |
| 體內(nèi)活性 | 每日處理0.5 mg/kg Tubastatin A 通過抑制HDAC6來促進(jìn)Tregs的抑制活性，在小鼠的炎癥與自身免疫模型中表現(xiàn)出效果，這些模型包括多種實(shí)驗(yàn)性結(jié)腸炎和完全主要組織相容性復(fù)合體（MHC）-不相容心臟異體移植的拒絕反應(yīng)。[2] |
| 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 : 3.7 mg/mL (10 mM), Heating is recommended.
H2O : 7.43 mg/mL (20 mM）, Sonication and heating are recommended.
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| 關(guān)鍵字 | Tubastatin A | Histone deacetylases | Inhibitor | Autophagy | inhibit | Apoptosis | HDAC | Tubastatin A Hydrochloride |
| 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 |
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