产品描述: | TPEN is a specific cell-permeable heavy metal chelator. |
靶点: |
Apoptosis;Autophagy;Reactive Oxygen Species;Apoptosis; ReactiveOxygenSpecies; Autophagy |
细胞实验: |
TPEN is dissolved in DMSO and then diluted with appropriate medium[1]. Human neuroblastoma cell line SH-SY5Y, are grown in Dulbecco's Modified Eagle's Medium (DMEM) mixed 1:1 with Ham's F-12 nutrient mixture containing 10% fetal bovine serum, 100 unit/mL penicillin and 100 μg/mL streptomycin at 37°C in a humidified 5% CO2 atmosphere. Two days before experimentation, cells are seeded at a density of 7×104 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 into the cells by using Calcium kit II fura-2. In brief, SH-SY5Ycells are incubated with 5 μM fura-2/AM in the presence of 0.04% Pluronic F-127, a dispersing agent to improve the efficiency of loading with fura-2, and 1.25 mM probenecid, a blocker of organic anion transport to prevent leakage of fura-2 from cells. After 1 hr incubation at 37°C, fura-2 fluorescence is measured at 500 nm emission after excitation at 340 nm (F340) or 380 nm (F380) using an Infinite M200 plate reader at 37°C. |
参考文献: |
1. Rahal ON, et al. Chk1 and DNA-PK mediate TPEN-induced DNA damage in a ROS dependent manner in human colon cancer cells. Cancer Biol Ther. 2016 Nov;17(11):1139-1148. 2. Ohkubo M, et al. Heavy metal chelator TPEN attenuates fura-2 fluorescence changes induced by cadmium, mercury and methylmercury. J Vet Med Sci. 2016 Jun 1;78(5):761-7. |
溶解性: |
Soluble in DMSO |
保存条件: |
-20℃ |
配置溶液浓度参考: |
|
1mg |
5mg |
10mg |
1 mM |
2.355 ml |
11.777 ml |
23.555 ml |
5 mM |
0.471 ml |
2.355 ml |
4.711 ml |
10 mM |
0.236 ml |
1.178 ml |
2.355 ml |
50 mM |
0.047 ml |
0.236 ml |
0.471 ml |
|
注意: |
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