Immunofluorescent analysis of HeLa cells using α-Tubulin (DM1A) Mouse mAb #3873 (red) and Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb #3377 (green). Blue = Hoechst 33342 (fluorescent DNA dye).
Flow cytometric analysis of Jurkat cells using Phospho-Histone H3 (Ser10) (D7N8E) XP® Rabbit mAb #53348 and Hoechst 33342 (DNA content). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
This product is soluble up to 50 mg/ml in deionized water or dimethylformamide yielding a clear yellow solution. Heating and/or sonication may be required. For standard fluorescence based assays, Hoechst 33342 stock solutions should be diluted in phosphate-buffered saline or mounting medium to a final working concentration of 1 μg/ml.
The lyophilized powder is stable for 1 year if kept below 0°C with desiccant. Aqueous solutions are stable for at least 1 month if kept in the dark between 2-8°C.
Hoechst 33342 (bisBenzimide H33342 trihydrochloride) is supplied as a lyophilized powder in 25 mg units. It can be used to examine cellular DNA in most fluorescent applications.
C27H28N6O 3(HCl) 3(H2O)
Hoechst 33342 is a specific fluorescent DNA stain that binds within the minor groove of double-stranded AT-rich regions. This stain can be used on both live and fixed cells (1). Replicating or dividing chromosomes will not stain with Hoechst 33342 DNA dye.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.