TPEN-TPEDA-DataSheet-生命科学试剂-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemETPENCat. No.: HY-100202CAS No.: 16858-02-9Synonyms: TPEDA分式: CHN分量: 424.54作靶点: Autophagy作通路: Autophagy储存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性数据体外实验 DMSO : 35.71 mg/mL (84.11 mM; Need ultrasonic)

2、Mass Solvent1 mg 5 mg 10 mg Concentration制备储备液1 mM 2.3555 mL 11.7775 mL 23.5549 mL5 mM 0.4711 mL 2.3555 mL 4.7110 mL10 mM 0.2355 mL 1.1777 mL 2.3555 mL请根据产品在不同溶剂中的溶解度，选择合适的溶剂配制储备液，并请注意储备液的保存式和期限。体内实验请根据您的实验动物和给药式选择适当的溶解案，配制前请先配制澄的储备液，再依次添加助溶剂(为保证实验结果的可靠性，体内实验的作液，建议您现现配，当天使；澄的储备液可以根据储存条件，适当保存；以下溶剂前的百

3、分指该溶剂在您配制终溶液中的体积占)：1. 请依序添加每种溶剂： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.08 mg/mL (4.90 mM); Clear solution2. 请依序添加每种溶剂： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.08 mg/mL (4.90 mM); Clear solution3. 请依序添加每种溶剂： 10% DMSO 90% corn oil1/3 Master of Small Molecules 您边的抑制剂师www.MedC

4、hemESolubility: 2.08 mg/mL (4.90 mM); Clear solutionBIOLOGICAL ACTIVITY物活性 TPEN种特异性的可渗透细胞的重 属螯合剂。体外研究 Heavy metal chelator TPEN attenuates fura-2 fluorescence changes induced by cadmium, mercury andmethylmercury. TPEN, a cell-permeable chelator for heavy metal cations with a low affinity for Ca2+. I

5、n cellsstimulated with 10 or 30 M cadmium chloride, the addition of TPEN at 3 hr after exposure significantlydecreases the elevated fura-2 fluorescence ratio to the basal levels within 10 min (119.62.4% or 1091.5%decrease in Ratio (F340/F380) induced by 10 or 30 M cadmium chloride, respectively), su

6、ggesting that acadmium chloride-induced increase in the fura-2 fluorescence ratio is dependent on an increase inintracellular heavy metal cations but not intracellular Ca2+ 1. TPEN is a metal chelator, which targets coloncancer cells through redox cycling of copper. TPEN reduces cell viability in a

7、dose- and time-dependentmanner. TPEN-induced cell death is also dependent on the redox cycling of copper since the copper chelatorneocuproine inhibited DNA damage and reduced pChk1, -H2AX, and ATM protein expression. Cell death bylow TPEN concentrations, involved ATM/ATR signaling in all 3 cell line

8、s, since pre-incubation with specificinhibitors of ATM and DNA-PK led to the recovery of cells from TPEN-induced DNA damage 2.PROTOCOLCell Assay 1 Human neuroblastoma cell line SH-SY5Y, are grown in Dulbeccos Modified Eagles Medium (DMEM) mixed1:1 with Hams F-12 nutrient mixture containing 10% fetal

9、 bovine serum, 100 unit/mL penicillin and 100 g/mL streptomycin at 37C in a humidified 5% CO2 atmosphere. Two days before experimentation, cells areseeded at a density of 7104 cells/cm2 in a 96-well plate. Cells in a 96-well plate are serum-starved for 4 hr;calcium indicator fura-2 is then loaded in

10、to the cells by using Calcium kit II fura-2. In brief, SH-SY5Ycells areincubated with 5 M fura-2/AM in the presence of 0.04% Pluronic F-127, a dispersing agent to improve theefficiency of loading with fura-2, and 1.25 mM probenecid, a blocker of organic anion transport to preventleakage of fura-2 fr

11、om cells. After 1 hr incubation at 37C, fura-2 fluorescence is measured at 500 nmemission after excitation at 340 nm (F340) or 380 nm (F380) using an Infinite M200 plate reader at 37C.Thechange in Ca2+i is reflected by the ratio of F340 and F380. To determine the changes in fura-2fluorescence ratio

12、induced by heavy metal compounds, cells are treated with manganese chloride, leadacetate, cadmium chloride , mercuric chloride and MeHg chloride dissolved in distilled water. We confirmedthat the cells adhered to the bottom of the plate after 6 hr exposure to heavy metal compounds. The cells arealso

13、 treated with three Ca2+ channel blockers, lanthanum chloride dissolved in distilled water, verapamil and2-APB dissolved in DMSO, 30 min before heavy metal exposure. The heavy metal chelator TPEN isdissolved in DMSO and added 3 hr after the stimulation with heavy metals to determine the contribution

14、 ofendogenous and exogenous heavy metals on fura-2 fluorescence changes. We measured the effect of TPEN(20 M) on the fura-2 fluorescence ratio after a 10 min treatment with TPEN, since our preliminaryexperiments showed that the effect of TPEN on fura-2 fluorescence reached maximum and stabilized wit

15、hin10 min of the treatment 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.2/3 Master of Small Molecules 您边的抑制剂师www.MedChemE Cancer Med. 2019 Apr 10.See more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Ohkubo M, et al. Heavy metal chelator TPEN attenuates fura-2 fluorescence changes induced by cadmium, mercury andmethylmercury. J Vet Med Sci. 2016 Jun 1;78(5):761-7.2. Rahal ON, et al. Chk1 and DNA-PK mediate TPEN-induced DNA damage in a ROS dependent manner in human colon cancer cells.Cancer