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MCE 国际站:PS48
CAS:1180676-32-7
品牌:MedChemExpress (MCE)
存储条件:Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
生物活性:PS48 是 一种 PDK1 激活剂,AC50 为 8 μM。
体外:PS48 activates full length PDK1 (His-PDK1- FL) and PDK1 50-359[Tyr288Gly;Gln292Ala] (His-PDK1 dm) with AC50s (the concentrations required to reach 50% of the maximal activation) of 7.95±0.2 and 10.0±2.0 μM, respectively. PS48 binds to PDK150–359 with a 1:1 stoichiometry and a binding affinity in the micromolar range (Kd=10.3 μM)[1]. PDK1 activator, PS48, has the ability to reverse the cell proliferation inhibition role of triptolide (TP) in vitro. The inhibition role of TP in cell number is significantly reversed by PS48. TP significantly increases the cell proportion in G0-G1 phase and decreases the cell proportion in G2-M and S phase. However, the effect of TP on cell cycle distribution is all reversed by PS48. In addition, suppression of PDK1/Akt/mTOR pathway by TP in high glucose (HG)-treated human renal mesangial cells (HRMCs) is also reversed by PS48, as well as the expression of Ki-67 and proliferating cell nuclear antigen (PCNA)[2]. MCE has not independently confirmed the accuracy of these methods. They are for reference only.
激酶试验:PDK1 活性测试使用 T308tide 作为 PDK1 的底物进行。简而言之,PDK1 活性测定是在室温 (22°C) 的 20 μL 混合物中进行,其中含有 50 mM Tris pH 7.5、0.05 mg/mL BSA、0.1% β-巯基乙醇、10 mM MgCl2、100 μM [γ32P]ATP (5-50 cpm/pmol)、0.003% Brij、150-500 ng PDK1 和 T308tide(0.1 至 1 mM)。适当时,PDK1 活性测定以 96 孔格式进行,使用 ep Motion 5070 将 4 μL 等分试样点在 p81 磷酸纤维素纸上,用 0.01% 磷酸洗涤,干燥,然后使用 PhosphoImager 技术进行曝光和分析。活性测量一式两份或一式三份进行,重复之间的差异小于 10%。实验至少重复两次[1]。
细胞试验:人肾系膜细胞 (HRMC) 使用含有 10% 胎牛血清的 1640 培养基在 37 °C、5% CO2 中培养。将细胞与正常 (5.5 mM) 或高 (25 mM) 浓度的 D-葡萄糖一起在无血清培养基中培养。 D-甘露醇 (25 mM) 用于控制渗透压。使用前将 TP 在 0.01% DMSO 中重新配制,并用培养基新鲜稀释至 10 ug/L。为了确定 PDK1 在 TP 增强的抗增殖中的具体作用,在 TP 治疗后应用 5 μM PS48(MedChem Express,美国)。 MTT法用于检测细胞增殖。 HRMC 以 1x105/mL 的密度接种到 96 孔板中。与不同化合物孵育 12、24、48 和 72 小时后,向每孔中添加 20 uL MTT (5 mg/mL)。然后将细胞再培养 2 小时,随后使用 DMSO(150 uL/孔)裂解。当甲臜晶体完全溶解时,在 570 nm 处测量光密度 (OD)。计算每组6个孔的算术平均OD[2]。
热销产品:Stigmasterol | L-Proline | Cyanine5 NHS ester (iodide) | Asundexian | L-Homocysteine | WZB117 | PF-573228 | Isochlorogenic acid A | PF-3644022 | Phthalic acid mono-2-ethylhexyl ester
研究领域:PI3K/Akt/mTOR
作用靶点:PDK-1
Trending products:Recombinant Proteins | Bioactive Screening Libraries | Natural Products | Fluorescent Dye | PROTAC | Isotope-Labeled Compounds | Oligonucleotides
参考文献:[1]. Hindie V, et al. Structure and allosteric effects of low-molecular-weight activators on the protein kinase PDK1. Nat Chem Biol. 2009 Oct;5(10):758-64.[2]. Han F, et al. Triptolide Suppresses Glomerular Mesangial Cell Proliferation in Diabetic Nephropathy Is Associated with Inhibition of PDK1/Akt/mTOR Pathway. Int J Biol Sci. 2017 Sep 21;13(10):1266-1275.
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