Hoechst 33342 analog trihydrochloride 

Hoechst 33342 analog trihydrochloride is a marker dye in Hoechst series. Hoechst is A live nuclear marker dye. Hoechst binds to the grooves in the DNA double strand, which tends to be A/ T-rich DNA strand. Although it binds to all nucleic acids, the A/ T-rich double strand DNA significantly enhances fluorescence intensity Therefore,Hoechst dye can be used for living cell labeling. The fluorescence intensity of Hoechst dye increases with the increase of pH of solution^[1]. Storage: Keep away from light.

Guide (The following is our recommended experimental protocol. This protocol is for reference only and should be adjusted according to your specific needs).
Preparation of Hoechst working solution
1.1 Preparation of stock solution
Prepare a 1 mg/mL Hoechst stock solution in DMSO.
Note: It is recommended to aliquot the Hoechst stock solution and store at -4°C or -20°C protected from light.
1.2 Preparation of working solution
Dilute the stock solution with pre-warmed serum-free cell culture medium or PBS to obtain a final concentration of 10 μg/mL Hoechst working solution.
Note: Adjust the concentration of Hoechst working solution according to actual needs and prepare fresh before use.
2. Cell staining (suspension cells)
2.1 Collect cells by centrifugation and wash twice with PBS, 5 min each time. Cell density: 1×10⁶/mL.
2.2 Add 1 mL Hoechst working solution and incubate at room temperature for 3-10 min.
2.3 Centrifuge at 400 g for 3-4 min and discard the supernatant.
2.4 Wash cells twice with PBS, 5 min each time.
2.5 Resuspend cells in 1 mL serum-free medium or PBS and observe using a fluorescence microscope or flow cytometer.
3. Cell staining (adherent cells)
3.1 Culture adherent cells on sterile coverslips.
3.2 Remove the coverslips from the culture medium and aspirate excess medium.
3.3 Add 100 μL dye working solution, gently shake to fully cover the cells, and incubate for 3-10 min.
3.4 Remove the dye working solution, wash 2-3 times with culture medium (5 min each), and observe using a fluorescence microscope or flow cytometer.


Notes
1. Adjust the concentration of Hoechst working solution according to actual needs.
2. This product is for research use only by professionals and is not intended for clinical diagnosis or treatment, nor for use in food or drugs.
3. For your safety and health, please wear a lab coat and disposable gloves during operation.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

699.97

C₃₂H₄₀Cl₅N₇

159277-19-7

ClCCN(CCCl)C1=CC=C(CCCC2=NC3=C(N2)C=C(C4=NC5=C(N4)C=C(N6CCN(CC6)C)C=C5)C=C3)C=C1.[H]Cl.[H]Cl.[H]Cl

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.

• [1]. Chazotte B. Labeling nuclear DNA with hoechst 33342. Cold Spring Harb Protoc. 2011 Jan 1;2011(1):pdb.prot5557.  [Content Brief]