|Powder: -20°C for 3 years | In solvent: -80°C for 1 year
PLX-4720 是一种有效且选择性的 B-Raf (V600E) 抑制剂,IC50为 13 nM,与 c-Raf-1(Y340D 和 Y341D 突变)同样有效。
规格 | 价格/CNY | 货期 | 数量 | |
---|---|---|---|---|
10 mg | ¥ 465 | 现货 | ||
50 mg | ¥ 997 | 现货 | ||
1 mL * 10 mM (in DMSO) | ¥ 371 | 现货 |
产品描述 | PLX-4720 is a potent and selective inhibitor of B-Raf (V600E) (IC50: 13 nM), equally potent to c-Raf-1(Y340D and Y341D mutations). |
靶点活性 | B-Raf (V600E):13 nM (cell free), B-Raf:160 nM (cell free), C-Raf1 (340D/Y341D):6.7 nM (cell free) |
体外活性 | PLX4720以13 nM的IC50抑制B-Raf(V600E)。与其高度选择性相符,PLX4720在携带B-Raf(V600E)的肿瘤细胞系中强效抑制ERK磷酸化,但对缺乏致癌性B-Raf的细胞不产生影响[1]。PLX4720处理显著提高了PTEN+细胞系中BIM表达,超过14倍,而在PTEN-细胞系中仅提高了四倍。在PTEN-细胞中,PLX4720与PI3K抑制剂的联合治疗,在mRNA和蛋白质水平上增强了BIM表达,并通过涉及AKT3和FOXO3a激活的机制增加了凋亡水平[2]。 |
体内活性 | 在B-Raf(V600E)-依赖性肿瘤异种移植模型中,口服PLX4720能显著延缓肿瘤生长,包括肿瘤退缩,且未观察到毒性证据[1]。在体内,8505c原位甲状腺肿瘤的PLX4720处理抑制了肿瘤的侵袭性,并显著上调了甲状腺分化标志物甲状腺转录因子1和配对盒基因8[3]。在肿瘤细胞植入后1周开始对原位甲状腺肿瘤进行PLX4720处理,能显著延迟肿瘤生长并减少远处转移,且未观察到毒性证据[4]。 |
激酶实验 | In vitro Raf kinase activities: The in vitro kinase activities of wild type Raf and mutants are determined by measuring phosphorylation of biotinylated-MEK protein using Perkin-Elmer's AlphaScreen Technology. For each enzyme (0.1 ng), 20-μL reactions are carried out in 20 mM Hepes (pH 7.0), 10 mM MgCl2, 1 mM DTT, 0.01% Tween-20, 100 nM biotin-MEK protein, various ATP concentrations, and increasing concentrations of PLX-4720 at room temperature. Reactions are stopped at 2, 5, 8, 10, 20, and 30 minutes with 5 μL of a solution containing 20 mM Hepes (pH 7.0), 200 mM NaCl, 80 mM EDTA, and 0.3% BSA. The stop solution also includes phospho-MEK Antibody, Streptavidin-coated Donor beads and Protein A Acceptor beads. The antibody and beads are preincubated in stop solution in the dark at room temperature for 30 minutes. The final dilution of antibody is 1/2,000, and the final concentration of each bead is 10 μg/mL. The assay plates are incubated at room temperature for one hour then are read on a PerkinElmer AlphaQuest reader [1]. |
细胞实验 | Cells are treated with various concentrations PLX-4720 for 24, 48, and 72 hours. Cell proliferation is measured by using the CellTiter-Glo Luminescent Cell Viability Assay or MTT assay. For cell cycle analysis, supernatant and cells are collected, pelleted, and fixed with 70% ethanol. Before staining with propidium iodide (10 μg/mL), cells are incubated for 1 hour at 37 °C in 0.5 mg/mL RNase I to rid samples of residual RNA contamination. Samples are then analyzed by using the EPICS XL apparatus. For the assessment of apoptosis, media and cells are harvested and pelleted before staining with annexin-FITC and propidium iodide. Samples are subsequently analyzed by using the EPICS XL apparatus [1]. |
动物实验 | Female athymic mice (NCr nu/nu) were implanted s.c. on day 0 with 30–60 mg COLO205 tumor fragments. Treatments began on day 11, when the mean estimated tumor mass was 104 mg (range, 95–113 mg). All animals were dosed with vehicle (5% DMSO, 1% methylcellulose) or PLX4720 suspended in vehicle by gavage daily for 14 consecutive days. Tumor burden (mg) was estimated from caliper measurements [1]. |
别名 | PLX4720 |
分子量 | 413.83 |
分子式 | C17H14ClF2N3O3S |
CAS No. | 918505-84-7 |
|Powder: -20°C for 3 years | In solvent: -80°C for 1 year
Ethanol: < 1 mg/mL (insoluble or slightly soluble)
DMSO: 60 mg/mL (144.99 mM)
H2O: < 1 mg/mL (insoluble or slightly soluble)
可选溶剂 | 浓度 体积 质量 | 1 mg | 5 mg | 10 mg | 25 mg |
DMSO | 1 mM | 2.4165 mL | 12.0823 mL | 24.1645 mL | 60.4113 mL |
5 mM | 0.4833 mL | 2.4165 mL | 4.8329 mL | 12.0823 mL | |
10 mM | 0.2416 mL | 1.2082 mL | 2.4165 mL | 6.0411 mL | |
20 mM | 0.1208 mL | 0.6041 mL | 1.2082 mL | 3.0206 mL | |
50 mM | 0.0483 mL | 0.2416 mL | 0.4833 mL | 1.2082 mL | |
100 mM | 0.0242 mL | 0.1208 mL | 0.2416 mL | 0.6041 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。
PLX-4720 918505-84-7 MAPK Raf Inhibitor inhibit Raf kinases PLX 4720 PLX4720 inhibitor