Powder: -20°C for 3 years | In solvent: -80°C for 1 year
Purmorphamine (Shh Signaling Antagonist VI) 是平滑受体激动剂,EC50为 1 μM。它阻止 BODIPY-cyclopamine 与 Smo 结合。它也是成骨细胞分化的诱导剂。
规格 | 价格/CNY | 货期 | 数量 | |
---|---|---|---|---|
1 mg | ¥ 396 | 现货 | ||
5 mg | ¥ 890 | 现货 | ||
10 mg | ¥ 1,250 | 现货 | ||
25 mg | ¥ 2,650 | 现货 | ||
50 mg | ¥ 3,450 | 现货 | ||
100 mg | ¥ 4,990 | 现货 | ||
1 mL * 10 mM (in DMSO) | ¥ 977 | 现货 |
产品描述 | Purmorphamine (Shh Signaling Antagonist VI), which directly binds and activates Smoothened, blocks BODIPY-cyclopamine binding to Smo. It also is an inducer of osteoblast differentiation. |
靶点活性 | Smo:1.5 μM |
体外活性 | 在大鼠中基于人间充质干细胞中,Purmorphamine 上调ALP表达. |
体内活性 | 在全能C3H10T1/2 细胞中(EC50=1 μM),Purmorphamine能够诱导骨生成。Purmorphamine 与Smoothened直接结合,而激活Hedgehog通路(IC50=1.5 μM)。 |
激酶实验 | Binding assay: Smo binding assays are conducted with BODIPY-cyclopamine and Smo-overexpressing cells as previously described4,5, using CMV promoter-based, SV40 origin-containing expression constructs for Smo-Myc3, the deletion mutant Smo?CRD (deletion of amino acids 68 to 182), and Smo?CT (deletion of amino acids 556 to 793). HEK 293T cells are grown on poly-D-lysine-treated glass coverslips in 12-well plates until 70% confluency and then transfected with the appropriate expression construct (0.5 ?g/well) using FuGene 6 according the manufacturer's protocols. Two days after transfection, the HEK 293T cells are incubated with DMEM containing 0.5% bovine calf serum, 5 nM BODIPY-cyclopamine, and varying concentrations of Purmorphamine (0, 1.5, or 5 ?M) (1 mL/well) for 1 h at 37 ℃. The Smo-overexpressing cells are then washed with 1 × PBS buffer (1 mL/well), mounted with DAPI-containing medium, and visualized using a Leica DM4500B fluorescence microscope. For binding assays using fixed cells, the Smo-overexpressing HEK 293T cells are fixed with 3% paraformaldehyde in 1 × PBS buffer for 10 min at room temperature (1 mL/well), treated with 1 × PBS containing 10 mM glycine and 0.2% sodium azide for 5 min (1 mL/well), washed with 1 × PBS buffer (1 mL/well), and treated with the Purmorphamine-containing media described above for 4 h at room temperature. |
细胞实验 | C3H10T1/2 cells are expanded in T175 flasks; cells at 13th passage are detached by trypsin/EDTA and diluted in the growth media. The resulting cell suspension is then plated into black clear bottom 384-well plates with 2500 cells/well in 100 μL growth medium using a Multi-dropTM liquid delivery system. After overnight incubation, cells attached to the bottom of the wells. A stock solution of each Purmorphamine in DMSO (500 nL) is delivered into corresponding well using a Mini TrakTM multiposition dispenser system to make a final concentration of 5 μM of Purmorphamine. Cells are then incubated at 37 ℃ with 5% CO2 in air atmosphere. After 4 days, the medium is removed and 10 μL of passive lysis buffer is added into each well. After 5 min, 10 μL of alkaline phosphatase substrate solution is added to each well. After incubating 15 min at room temperature, the plates are read on an Acquest high-throughput plate reader following the manufacturer's protocol.(Only for Reference) |
别名 | Shh Signaling Antagonist VI |
分子量 | 520.62 |
分子式 | C31H32N6O2 |
CAS No. | 483367-10-8 |
Powder: -20°C for 3 years | In solvent: -80°C for 1 year
DMSO: 52.1 mg/mL (100 mM)
可选溶剂 | 浓度 体积 质量 | 1 mg | 5 mg | 10 mg | 25 mg |
DMSO | 1 mM | 1.9208 mL | 9.6039 mL | 19.2079 mL | 48.0197 mL |
5 mM | 0.3842 mL | 1.9208 mL | 3.8416 mL | 9.6039 mL | |
10 mM | 0.1921 mL | 0.9604 mL | 1.9208 mL | 4.802 mL | |
20 mM | 0.096 mL | 0.4802 mL | 0.9604 mL | 2.401 mL | |
50 mM | 0.0384 mL | 0.1921 mL | 0.3842 mL | 0.9604 mL | |
100 mM | 0.0192 mL | 0.096 mL | 0.1921 mL | 0.4802 mL |
对于不同动物的给药剂量换算,您也可以参考 更多...
请在以下方框中输入您的动物实验信息后点击计算,可以得到母液配置方法和体内配方的制备方法: 比如您的给药剂量是10 mg/kg,每只动物体重20 g,给药体积100 μL,一共给药动物10 只,您使用的配方为5% DMSO+30% PEG300+5% Tween 80+60% ddH2O。那么您的工作液浓度为2 mg/mL。
母液配置方法:2 mg 药物溶于 50 μL DMSO (母液浓度为 40 mg/mL), 如您需要配置的浓度超过该产品的溶解度,请先与我们联系。
体内配方的制备方法:取 50 μL DMSO 主液,加入 300 μL PEG300, 混匀澄清,再加 50 μL Tween 80,混匀澄清,再加 600 μL ddH2O, 混匀澄清。
您可能有的问题的答案可以在抑制剂处理说明中找到,包括如何准备库存溶液,如何存储产品,以及基于细胞的分析和动物实验需要特别注意的问题。
Purmorphamine 483367-10-8 Autophagy GPCR/G Protein Stem Cells Hedgehog/Smoothened Smo inhibit Smoothened Inhibitor Shh Signaling Antagonist VI inhibitor