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CAS No. : 23491-45-4
MCE 国际站:Hoechst 33258 trihydrochloride
产品活性:Hoechst 33258 trihydrochloride 是 Hoechst 系列的标记染料。Hoechst 系列是活细胞核标记染料。Hoechst 通过结合 DNA 双链中的小沟而结合核酸。Hoechst 倾向于结合富含 A/T 的 DNA 链。同时,富含 A/T 的双链 DNA 使荧光强度显著增强。Hoechst 可穿过细胞膜,结合活细胞或固定细胞。Hoechst 染料的荧光强度随着溶液 pH 升高而增强。
研究领域:Others
作用靶点:Fluorescent Dye
In Vitro: Preparation of Hoechst working solution
1.1 Preparation of stock solution
Take 10 mg and dissolve in 5 mL DMSO
Note: It is recommended that the stock solution be stored at 4℃ or - Store at 20℃ in the dark and avoid repeated freezing and thawing.
1.2 Preparation of Hoechst working solution
The diluted stock solution was dissolved in serum-free cell culture medium or PBS to obtain Hoechst working solution with a final concentration of 10 μg/mL.
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 for 3-5 min at 4℃ and discard the supernatant. Wash 2 times with PBS, 5 minutes each. The cell density was 1×106/mL.
b. Add 1 mL of working solution and incubate at room temperature for 3-10 minutes.
c. Centrifuge at 400 g for 3-4 minutes at 4℃ and discard the supernatant.
d. Wash 2 times with PBS, 5 min each.
e. Resuspend cells in serum-free cell culture medium or PBS. Fluorescence microscopy or flow cytometry observation.
2.2 Adherent cells
a. Adherent cells were cultured on sterile coverslips.
b. Remove the coverslip from the medium and aspirate excess medium. c. Add 100 μL of working solution, shake gently to completely cover the cells, and incubate at room temperature for 3-10 minutes.
d. Wash 2 times with medium, 5 min each. Fluorescence microscope or flow cytometry observation.
Store
4°C, 1 year. Avoid light
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 medicine, household or other purposes.
3. For your safety and health, please wear a lab coat and disposable gloves for operation.
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