1. Hoechst 33258 Staining Solution (Ready-To-Use) 10 ml

Product Information

Description

The molecular formula of Hoechst 33258 is C25H24N6O·3HCl, the molecular weight is 533.88, and the CAS number is 23491-45-4. Hoechst 33258 is a blue fluorescent dye that can penetrate cell membrane, and its fluorescence is significantly enhanced when combined with double-stranded DNA. The maximum excitation wavelength is 346 nm, and the maximum emission wavelength is 460 nm. Hoechst 33258 is commonly used for apoptosis detection, nuclear staining, or routine DNA staining. It can be used to stain the nuclei of fixed cells and tissues or non-fixed cells and tissues. After staining, it can be observed by fluorescence microscope or detected by flow cytometry. When observed by fluorescence microscope, it is excited by ultraviolet light, which is blue fluorescence.

This product is a ready-to-use solution, the concentration of which is optimized to meet the needs of all kinds of routine staining.

Storage and Handling Conditions

Wet ice transportation; Store at 4℃ away from light, valid for 6 months.

Component

Assay Protocol

1. For fixed cell or tissue sections:

a. For the fixed cell or tissue samples, remove the fixing solution after fixation, and wash with PBS or another appropriate buffer 2-3 times, 3-5 min each time (for the detection of suspended cells, please follow the routine operation method of suspended cells, and centrifugation and other steps shall be added in all experiments);

b. (Optional step) If the fixed cells or tissues need to be treated with immunofluorescence staining, the relevant treatment should be performed first, and then Hoechst 33258 staining should be performed after the treatment;

c. Directly take an appropriate amount of the product Hoechst 33258 dye solution (ready-to-use type) to cover the sample, and incubate at room temperature for about 5 min in the dark;

d. Remove the dye solution and wash it with PBS or another appropriate buffer 2-3 times, 3-5 min each time;

e. After sealing the plates or directly placed under a fluorescence microscope, the excitation wavelength was 350 nm, and the emission wavelength was 460 nm.

2. For living cells or cultured tissues:

a. Add an appropriate amount of Hoechst 33258 dye solution (ready-to-use type) to the cell culture to fully cover the sample to be stained; generally, 1 mL of staining solution should be added to a well of a 6-well plate, and 100 μL of staining solution should be added to a well of a 96-well plate. The samples were incubated for 10-20 min in an incubator at 37℃.

b. Remove the dye solution and wash it with PBS or another appropriate buffer 2-3 times, 3-5 min each time;

c. Directly observed under a fluorescence microscope, the excitation wavelength was 350 nm, and the emission wavelength was 460 nm.

Note:

1. All fluorescent dyes have the problem of fluorescence quenching. It is recommended to take photos as soon as possible after dyeing. To slow down fluorescence quench, an anti-fluorescence quench sealer (G1401 recommended) can be used.

2. Wear a lab coat and disposable gloves during operation.

For Research Use Only!