Torin 1
作者：齐岳
发布时间：2023-09-14
08:57:46
点击数：
产品名称：Torin 1
产品描述：
| 产品描述 | Torin 1 is an effective inhibitor of mTORC1/2 with (IC50: 2 nM/10 nM); has 1000-fold selectivity for mTOR than PI3K. |
| 靶点活性 | mTORC2:10 nM, mTORC1:2 nM |
| 体外活性 | Torin1（20 mg/kg）对U87 mg异种移植模型有效的,且可有效抑制肿瘤和外周组织中的mTOR下游效应蛋白. |
| 体内活性 | Torin1通过耐雷帕霉素机制引起细胞周期阻滞,并且其不依赖于mTORC2。Torin1比雷帕霉素对mTORC1依赖的表现型干扰更完全。在近期的一项研究中表明,Torin1通过活化人内分泌细胞系BON 中MEK/ERK/c-Jun通路,能够增加神经降压素分泌和基因表达。Torin1（2、10 nM）可分别抑制mTORC1 和mTORC2底物的磷酸化。 |
| 激酶实验 | mTORC1 and mTORC2 in Vitro Kinase Assays: To produce soluble mTORC1, HEK-293T cell lines that stably express N-terminally FLAG-tagged Raptor are generated using vesicular stomatitis virus G-pseudotyped MSCV retrovirus. For mTORC2, similar HeLa cells that stably express N-terminally FLAG-tagged Protor-1 are generated. Both complexes are purified by lysing cells in 50 mm HEPES, pH 7.4, 10 mm sodium pyrophosphate, 10 mm sodium β-glycerophosphate, 100 mm NaCl, 2 mm EDTA, 0.3% CHAPS. Cells are lysed at 4 °C for 30 min, and the insoluble fraction is removed by microcentrifugation at 13,000 rpm for 10 min. Supernatants are incubated with FLAG-M2 monoclonal antibody-agarose for 1 h and then washed three times with lysis buffer and once with lysis buffer containing a final concentration of 0.5 M NaCl. Purified mTORC1 is eluted with 100 μg/mL 3×FLAG peptide in 50 mm HEPES, pH 7.4, 100 mm NaCl. Eluate can be aliquoted and stored at -80 °C. Substrates S6K1 and Akt1 are purified. Kinase assays are performed for 20 min at 30 °C in a final volume of 20 μL consisting of the kinase buffer (25 mm HEPES, pH 7.4, 50 mm KCl, 10 mm MgCl2, 500 μm ATP) and 150 ng of inactive S6K1 or Akt1 as substrates. Reactions are stopped by the addition of 80 μL of sample buffer and boiled for 5 min. Samples are subsequently analyzed by SDS-PAGE and immunoblotting. |
| 细胞实验 | Cell viability is assessed with the CellTiter-Glo Luminescent Cell Viability Assay. On Day 0, 96-well plates are seeded with 500 cells per well and grown overnight. On Day 1, cells are treated with the appropriate compounds and subsequently analyzed on Days 3-5. For analysis, plates are incubated for 60 min at room temperature; 50 μL of CellTiter-Glo reagent is added to each well, and plates are mixed on an orbital shaker for 12 min. Luminescence is quantified on a standard plate luminometer. (Only for Reference) |
| 分子量 | 607.62 |
| 分子式 | C35H28F3N5O2 |
| CAS No. | 1222998-36-8 |
存储
Powder: -20°C for 3 years | In solvent: -80°C for 2 years
溶解度
DMSO: 2.5 mM
( < 1 mg/mL refers to the product slightly soluble or insoluble )
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