Hoechst 33342 Staining Solution (1 mg/mL), 1 mL

Product Information

Description

Hoechst 33342 (bisBenzimide H 33342, HOE 33342), the molecular formula is C₂₇H₂₈N₆O·3HCl, molecular weight is 561.93, CAS number is 23491-52-3. It is a blue fluorescent dye that can penetrate the cell membrane to stain DNA in the nucleus. This dye has weak fluorescence in the solution, but the fluorescence becomes brighter when combined with the small groove of the DNA in the AT-enriched area, so it is used for ordinary nuclear staining or conventional DNA staining.

This product is provided as a solution with a concentration of 1 mg/mL of Hoechst 33342. When used for staining, the usual working concentration is 0.5-10 μg/mL. Hoechst 33342 has a maximum excitation wavelength of 346 nm and a maximum emission wavelength of 460 nm; After being combined with double-stranded DNA, the maximum excitation wavelength is 350 nm and the maximum emission wavelength is 461 nm. It can be used to label the nucleus of cells or tissues under fixed or non-fixed conditions, and can be detected by fluorescence microscope or flow cytometer and other instruments.

Storage and Handling Conditions

Ship with wet ice；store at -20°C away from light, valid within 12 months。

Component

Procedures

1. The Hoechst 33342 staining solution (1 mg/mL) was diluted to 0.5-10 μg/mL Hoechst 33342 staining working solution with PBS or other suitable buffer;

2. For fixed cell and tissue samples:

a. For fixed cell or tissue samples, remove the fixative after fixation and wash 2-3 times with PBS or other suitable buffer，3-5 min each time (if detecting suspension cells, please follow the routine operation of suspension cells, and add steps such as centrifugation for all experiments);

b. (optional) If the fixed cells or tissues need to be processed for immunofluorescence staining, etc., the relevant processing is prioritized, and the Hoechst 33342 staining is performed after the processing;

c. Take an appropriate amount of Hoechst 33342 staining working solution to stain the fixed cells or tissues, cover the sample and incubate at room temperature for about 5 minutes;

d. After removing the Hoechst 33342 staining working solution, wash with PBS or other suitable buffer 2-3 times for 3-5 min each time;

e. Sealing or observe it under a fluorescence microscope directly,Ex/Em: 350 nm/460 nm.

3. For living cells or cultured tissues:

a. Add an appropriate amount of Hoechst 33342 staining working solution to the sample to be stained. The sample must be fully covered. Incubate in a 37°C incubator for 10-20 min. Usually 1 mL of staining solution is added to one well of a six-well plate，100 μL of staining solution should be added to one well of the 96-well plate；

b. Remove the staining solution,wash 2-3 times with PBS or other suitable buffer, 3-5 min each time;

c. Observe it under a fluorescence microscope directly,Ex/Em: 350 nm/460 nm.

Note

1. The incubation concentration and time of Hoechst 33342 staining working solution can be adjusted according to the actual situation.

2. Fluorescent dyes have quenching problems, try to complete the detection on the same day after dyeing; use anti-fluorescence quenching mounting tablets (recommended Servicebio G1401) to slow down fluorescence quenching.

3. For your health and safety, please wear lab coats and gloves during operation.

For Research Only! 

https://www.servicebio.com/goodsdetail?id=5876