化合物CP466722 T6261

Product Introduction
Bioactivity


英文名: CP-466722

描述: CP-466722 是一种有效且可逆的 ATM 抑制剂，IC50值为 4.1 μM。它不会抑制细胞中的 ATR 和 PI3K 或 PIKK 家族成员。

细胞实验: HeLa or A-T (GM02052 expressing hTERT) cells are plated in triplicate and incubated for 24 hours. Cells are pre-treated: DMSO, CP466722 or KU55933 prior to IR (0-10 gy). Cells are incubated for 4 hours following IR before media is removed, cells washed (PBS), trypsined, counted and re-plated (2000 cells/plate, 10 cm plates) in the absence of drug and incubated for 10 days. Prior to colony counting, cells are washed (PBS), stained (PBS, 0.0037%v/v-formaldehyde, 0.1%w/v-crystal violet), rinsed (dWater) and dried. Defined populations (>50 cells) are counted as one surviving colony, data are calculated as percentage surviving colonies relative to control plates +/− SE. (Only for Reference)

激酶实验: In vitro kinase assays [1] : To screen for small molecule inhibitors of ATM kinase activity, an in vitro kinase assay is adapted, and an ELISA assay develops which measured the phosphorylation status of the ATM downstream target p53. Recombinant GST-p53(1-101) and full-length Flag-tagged ATM & ATR are purified for use in the ELISA and in vitro kinase assays. Briefly, Nunc 96 well Maxisorp plates are coated overnight (4 °C) with 2μg of purified, recombinant GST-p53(1-101) in PBS. All subsequent incubations are performed at room temperature. The plates are washed (0.05%v/v-Tween/PBS) before addition of purified recombinant full-length ATM kinase (30 ng–60 ng) in a final volume of 80μL of reaction buffer (20 mM HEPES, 50 mM NaCl, 10 mM MgCl2, 10 mM MnCl2, 1 mM DTT and 1 μM ATP) in the presence or absence of CP-466722. CP-466722 (10 μM) is added to plates in duplicate and the kinase assay is incubated (90 minutes). Plates are washed (0.05%v/v-Tween/PBS), blocked (1hour, 1%w/v-BSA/PBS) and rinsed before anti-Phospho(Ser15)-p53 antibody (1:1000/PBS) is added to the plates and incubated (1hour). To reduce non-specific binding plates are washed (0.05%v/v-Tween/PBS) prior to incubation (1hour) with HRP-conjugated goat anti-rabbit IgG secondary antibody (1:5000/PBS). Secondary antibody that is linked to the phosphorylated GST-p53(1–101) protein is detected with TMB substrate reagent. Plates are developed (15 minutes–30 minutes) and the reaction is stopped (1 M H2SO4 final concentration) before absorbance is determined (λ450 nM). CP-466722 that inhibits ATM kinase activity in ELISA assays, are characterized with respect to inhibition of ATM/ATR kinases using in vitro kinase assays. Western blotting using the anti-Phospho(Ser15)-p53 antibody is used as a readout of ATM/ATR inhibition. Extended analysis of CP466722 (10 μM) against a commercially available panel of kinases is performed by Upstate.

体外活性: CP-466722 is used as a potential inhibitor to inhibits the activity of ATM kinase, which phosphorylates GST-p53(1–101) substrate. CP-466722 also exhibits the decreasing activities against abl and src kinases. In MCF-7 human breast cancer cells and primary and immortalized diploid human fibroblasts, CP-466722 suppress ATM-dependent p53 induction. In HeLa cells, CP-466722(6 μM), leads to the inhibition in ATM-dependent phosphorylation by reversibly inhibiting ionizing radiation (IR)-induced ATM kinase activity.

存储条件: Powder: -20°C for 3 years | In solvent: -80°C for 1 year

溶解度: DMSO : 7 mg/mL (20.04 mM)


关键字: ATM and RAD3 related | CP-466722 | Inhibitor | Ataxia telangiectasia mutated | ATM/ATR | inhibit

相关产品: Schisandrin B | ATR-IN-29 | Torin 2 | VE-821 | KU-55933 | AZ31 | ETP-46464 | Ceralasertib | Gartisertib | Wortmannin

相关库: HIF-1 Signaling Pathway Compound Library | Anti-Prostate Cancer Compound Library | ReFRAME Related Library | Anti-Cancer Metabolism Compound Library | Cancer Cell Differentiation Compound Library | Kinase Inhibitor Library | Inhibitor Library | Anti-Cancer Active Compound Library | Target-Focused Phenotypic Screening Library | Reprogramming Compound Library

化合物CP466722 T6261信息由TargetMol中国为您提供，如您想了解更多关于化合物CP466722 T6261报价、型号、参数等信息，欢迎来电或留言咨询。

注：该产品未在中华人民共和国食品药品监督管理部门申请医疗器械注册和备案，不可用于临床诊断或治疗等相关用途