| 名稱 | HSF1A |
| 描述 | HSF1A is a cell-permeable activator of HSF1 that protects mammalian cells against stress-induced apoptosis. |
| 細(xì)胞實(shí)驗(yàn) | PC12 cells seeded into a 96-well plate (5×104 cells/well) are treated with increasing concentrations of HSF1A (2, 4, 8 and 12 μM) for 15 h, at which time httQ74-GFP expression is stimulated by incubation in the presence of 1 μg/mL Doxycycline for 5 d. Cell viability is assessed via the XTT viability assay. |
| 激酶實(shí)驗(yàn) | Protein extracts are generated from mammalian, yeast and E. coli cultures using biotin-binding buffer (20 mM HEPES, 5 mM MgCl2, 1 mM EDTA, 100 mM KCl, 0.03% NP-40) supplemented with 1% Trition-X100 and protease inhibitors. Approximately 0.5 mg of protein extract is incubated with 100 μM HSF1A-Biotin for 4 h at 4°C and HSF1A-Biotin associated proteins captured by with NeutrAvidin Agarose Resin. After washing in biotin binding buffer proteins are eluted using 50 μL biotin elution buffer (100 mM Tris, 150 mM NaCl, 0.1 mM EDTA, 2 mM D-biotin), resolved on a 4-20% SDS-PAGE, and immunoblotted. For purified TRiC and Hsp70 analyses, 5 nM protein is incubated in biotin-binding buffer+0.5% Triton X-100 with 100 μM biotin or 100 μM HSF1A-Biotin for 4 h at 4°C and captured with NeutrAvidin Resin. For NiNTA purified yeast Tcp1, different concentrations of Tcp1 0.5 μM, 1 mM, 2 mM, 3 mM and 4 mM in 25 mM Hepes pH 7.5, 150 mM NaCl are incubated with 0.5 μM Biotin or HSF1A-Biotin for 4 h at 4°C and captured with NeutrAvidin Resin. |
| 動(dòng)物實(shí)驗(yàn) | RatTen-week-old Wistar Kyoto rats (WKY) are used. The rats are housed at a constant temperature (22°C) on a 12-h light/dark cycle with food and tap water. The animals are arranged into three groups: WKY rats (the control group), DOX rats and DOX rats treated with HSF1A. Each group contain five animals. The DOX group is injected with DOX (5?mg/kg) for 6 consecutive weeks intraperitoneal injection to achieve a cumulative dose of 30?mg/kg, which has been well documented to achieve cardiotoxicity. The small molecular HSF1 activator HSF1A (100?mg/kg/day) is injected intraperitoneally. |
| 體外活性 | HSF1A是一種小分子HSF1激活劑，能保護(hù)細(xì)胞免受應(yīng)激誘導(dǎo)的凋亡，抑制TRiC活性，但不擾亂ATP水解。TRiC復(fù)合體的遺傳失活或耗盡會(huì)導(dǎo)致人類HSF1激活，而HSF1A可抑制純化的TRiC與HSF1之間的直接相互作用。此外，使用與HSF1A偶聯(lián)的FITC進(jìn)行的熒光偏振實(shí)驗(yàn)顯示，HSF1A-FITC以約600 nM的親和力結(jié)合到TRiC的純化Tcp1亞基。通過將純化的Tcp1滴定到含有500 nM生物素或HSF1A-生物素的結(jié)合反應(yīng)中，這一點(diǎn)得到定性驗(yàn)證。通過統(tǒng)計(jì)含有聚集體的細(xì)胞數(shù)占總細(xì)胞數(shù)的比例進(jìn)行量化，顯示在HSF1A濃度低至2 μM時(shí)，觀察到含聚集體細(xì)胞的數(shù)量減少。預(yù)處理12 μM HSF1A的細(xì)胞，約20％顯示出通過熒光顯微鏡可見的聚集體，且隨劑量增加細(xì)胞中含聚集體的比例持續(xù)減少。 |
| 體內(nèi)活性 | HSF1A通過增強(qiáng)HSF1活性、穩(wěn)定HSF1表達(dá)并減輕多柔比星(DOX)引起的心臟損傷效果顯著。在多柔比星（累積劑量：30mg/kgw）和多柔比星聯(lián)合HSF1A（100mg/kgw/天）挑戰(zhàn)的WKY大鼠中，補(bǔ)充HSF1A能顯著提升心臟功能至對(duì)照組水平。研究表明，HSF1A可促進(jìn)人HSF1向核內(nèi)轉(zhuǎn)移，提高蛋白質(zhì)伴侶表達(dá)量，改善蛋白質(zhì)錯(cuò)誤折疊及細(xì)胞死亡在神經(jīng)退行性疾病模型中的情況。超聲心動(dòng)圖結(jié)果顯示，HSF1A亦能緩解DOX引起的心臟功能障礙。 |
| 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 : 50 mg/mL (122.09 mM)
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| 關(guān)鍵字 | Inhibitor | HSP | inhibit | Heat shock proteins | HSF1A | HSF-1A |
| 相關(guān)產(chǎn)品 | SNX2112 | Teprenone | SNX0723 | Gedunin | Tamoxifen Citrate | Rifabutin | Paeoniflorin | Tamoxifen | Ethoxyquin | Ganetespib | DN401 | KRIBB11 |
| 相關(guān)庫 | 抑制劑庫 | 抗乳腺癌化合物庫 | 經(jīng)典已知活性庫 | 抗癌化合物庫 | 已知活性化合物庫 | 轉(zhuǎn)錄因子庫 | 細(xì)胞凋亡化合物庫 | 細(xì)胞骨架化合物庫 | NO PAINS 化合物庫 |