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CAS No. : 1003197-00-9
MCE 国际站:MitoSOX Red
产品活性:MitoSOX Red 是一种特异性靶向活细胞线粒体的新型荧光探针,具有细胞膜渗透性。MitoSOX Red 进入线粒体后会被超氧化物氧化,而不会被其他 ROS 或 RNS 生成系统氧化。MitoSOX Red 随后与线粒体内核酸结合,产生强红色荧光。MitoSOX Red 可以作为荧光指示剂,特异性检测超氧化物。另外,超氧化物歧化酶 (Superoxide dismutase,SOD) 能够预防 MitoSOX Red 氧化。
激发/发射波长:396/610 nm。
研究领域:Others | Immunology/Inflammation | NF-κB | Metabolic Enzyme/Protease
作用靶点:Fluorescent Dye | Reactive Oxygen Species | Mitochondrial Metabolism
In Vitro: General Protocol
1. Preparation of MitoSOX Red working solution
1.1 Preparation of the stock solution
Dissolve 50 μg MitoSOX Red in 13 μL DMSO to obtain 5 mM of stock solution.
Note: It is recommended to store the stock solution at -20°C and -80°C away from light and avoid repetitive freeze-thaw cycles.
1.2 Preparation of MitoSOX Red working solution
Dilute the stock solution in serum-free cell culture medium or PBS to obtain 1-10 μM of working solution.
Note: Please adjust the concentration of MitoSOX Red working solution according to the actual situation.
2. Cell staining
2.1 Suspension cells(6-well plate)
a. Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.The cell density is 1×106/mL.
b. Add 1 mL of working solution, and then incubate at room temperature for 5-30 minutes.
c. Centrifuge at 400 g at 4°C for 3-4 minutes and then discard the supernatant.
d. Wash twice with PBS, 5 minutes each time.
e. Resuspend cells with serum-free cell culture medium or PBS. Observation by fluorescence microscopy or flow cytometry.
2.2 Adherent cells
a. Culture adherent cells on sterile coverslips.
b. Remove the coverslip from the medium and aspirate excess medium.
c. Add 100 μL of working solution, gently shake it to completely cover the cells,and then incubate at room temperature for 5-30 minutes.
d. Wash twice with medium, 5 minutes each time. Observation by fluorescence microscopy or flow cytometry.
Storage
-20°C, 1 year. Protect from light
Precautions
1. Please adjust the concentration of MitoSOX Red working solution according to the actual situation.
2. This product is for R&D use only, not for drug, household, or other uses.
3. For your safety and health, please wear a lab coat and disposable gloves to operate.
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