Hoechst 34580 (HOE 34580), a cell-permeable fluorescent dye, is used for staining DNA and nuclei.

Hoechst 34580 is a good candidate for treating Alzheimer's disease by inhibiting Aβ formation. 50 μM Aβ42 solutions co-incubated with 100, 25, 12.5, 3.125, 0.78, and 0.1, 0.01 μM Hoechst 34580 at 37 °C for 70 h. Hoechst 34580 can inhibit the aggregation of Aβ42 in a dose-dependent manner. The IC50 is obtained by measuring the concentration of Hoechst 34580 while maintaining the Aβ42 concentration which gave 0.86±0.05 μM for Hoechst 34580.

Instructions:
I. Solution preparation
1. Preparation of stock solution: Prepare 1 mg/mL Hoechst 34580 stock solution with DMSO.
Note: It is recommended to store Hoechst stock solution at -4℃ or -20℃ in the dark after aliquoting.
2. Preparation of working solution: Dilute the stock solution with pure DMEM or PBS before use, and the final concentration is 10 μg/mL Hoechst working solution.
Note: Please adjust the concentration of Hoechst working solution according to actual conditions and prepare it before use.
II. Cell staining (suspended cells)
1. Collect cells by centrifugation and wash twice with PBS for 5 minutes each. The cell density is 1×10^6/mL
2. Add 1 mL Hoechst working solution and incubate at room temperature for 3-10 minutes.
3. Centrifuge at 400 g for 3-4 minutes and discard the supernatant.
4. Add PBS to wash cells twice for 5 minutes each.
5. Resuspend the cells with 1 mL serum-free medium or PBS, and observe using a fluorescence microscope or flow cytometer.
III. Cell staining (adherent cells)
1. Culture the adherent cells on a sterile coverslip.
2. Remove the coverslip from the culture medium and remove the excess culture medium.
3. Add 100 μL of dye working solution, gently shake to completely cover the cells, and incubate for 3-10 minutes.
4. Aspirate the dye working solution, wash with culture medium 2-3 times, 5 minutes each time, and observe using a fluorescence microscope or flow cytometer.
Precautions
1. Please adjust the concentration of Hoechst working solution according to your actual situation.
2. This product is limited to scientific research by professionals and shall not be used for clinical diagnosis or treatment, nor for food or medicine.
3. For your safety and health, please wear a lab coat and disposable gloves when operating.