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CAS No. : 23491-52-3
MCE 國(guó)際站:Hoechst 33342
產(chǎn)品活性:Hoechst 33342 是 Hoechst 系列的標(biāo)記染料,。Hoechst 系列是活細(xì)胞核標(biāo)記染料。Hoechst 通過結(jié)合 DNA 雙鏈中的小溝而結(jié)合核酸,。Hoechst 傾向于結(jié)合富含 A/T 的 DNA 鏈,。同時(shí),富含 A/T 的雙鏈 DNA 使熒光強(qiáng)度顯著增強(qiáng),。Hoechst 可穿過細(xì)胞膜,,結(jié)合活細(xì)胞或固定細(xì)胞。Hoechst 染料的熒光強(qiáng)度隨著溶液 pH 升高而增強(qiáng),。
研究領(lǐng)域:Autophagy
作用靶點(diǎn):Autophagy
In Vitro: General Protocol
Preparation of Hoechst working solution
1.1 Preparation of the stock solution
Dissolve 10 mg of in 5 mL DMSO
Note: It is recommended to store the stock solution at 4℃ or -20℃ away from light and avoid repetitive freeze-thaw cycles.
1.2 Preparation of Hoechst working solution
Dilute the stock solution in serum-free cell culture medium or PBS to obtain final concentration 10 μg/mL Hoechst working solution.
Note: Please adjust the concentration of Hoechst working solution according to the actual situation.
1.Cell staining
2.1 Suspension cells(6-well plate)
a. Centrifuge at 1000 g at 4℃ 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 3-10 minutes.
c. Centrifuge at 400 g at 4℃ 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 3-10 minutes.
d. Wash twice with medium, 5 minutes each time.Observation by fluorescence microscopy or flow cytometry.
Precautions
1. Please adjust the concentration of Hoechst 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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