Solarbio Hoechst 33342 Stain Solution 10ug/ml (ready-to-use)

Product Code: C0030

Specification: 10ml/50ml

Storage: Store in the dark at -20°C, valid for one year.

Product Introduction:

• Hoechst 33342, also known as bisBenzimide H 33342 or HOE33342, is a blue fluorescent dye capable of penetrating cell membranes with low toxicity to cells.
• It is commonly used for detecting cell apoptosis, and after staining, observations can be made using fluorescence microscopy or flow cytometry.
• Hoechst 33342 is also used for nuclear staining or routine DNA staining. Its maximum excitation wavelength is 346nm, and the maximum emission wavelength is 460nm. When bound to double-stranded DNA, the excitation wavelength shifts to 350nm and the emission wavelength to 461nm.
• This Hoechst 33342 staining solution is ready-to-use and suitable for nuclear staining of fixed or live cells.

Instructions for Use:

For fixed cells or tissues:

a. After fixation, wash to remove the fixative. If immunofluorescence staining is required, perform it first before Hoechst 33342 staining. If no other staining is needed, proceed directly to Hoechst 33342 staining.

b. For adherent cells or tissue sections, add a small amount of Hoechst 33342 staining solution to cover the sample. For suspended cells, add three times the volume of the sample to be stained, mix well, and incubate at room temperature for 3-5 minutes.

c. Remove the staining solution, and wash 2-3 times with TBST, PBS, or physiological saline for 3-5 minutes each time.

d. Observe directly under a fluorescence microscope or after sealing the slide. Apoptotic cell nuclei appear densely stained or fragmented.

For live cells or tissues:

a. Add an appropriate amount of Hoechst 33342 staining solution, ensuring full coverage of the sample to be stained. Add 1 ml for each well of a six-well plate or 100μl for each well of a 96-well plate.

b. Incubate at an appropriate temperature for cell culture for 20-30 minutes. Discard the solution, wash 2-3 times with PBS or culture medium, and then proceed to fluorescence detection.

Precautions:

Fluorescent dyes are prone to quenching; it is recommended to use an anti-fade mounting medium. Try to complete the detection on the same day after staining, and observe live cells or tissues immediately after staining. For your safety and health, please wear laboratory attire and disposable gloves during the operation.

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