Hoechst 33342, blue fluorescent nucleic acid stain

Cat. # Quantity Price Lead time
2G010 5 mL, 10 mg/mL in Water $85 in stock
1H010 100 mg $125 in stock
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Hoechst 33342 (bisbenzimide, HOE 33342) is a cell-permeable blue-emitting fluorescent dye that binds strongly to adenine-thymine-rich regions in the minor groove of double-stranded DNA. Although Hoechst 33342 can bind to all nucleic acids, AT-rich dsDNA strands enhance its fluorescence considerably.

Hoechst 33342 bound with DNA has excitation/emission maxima at 351/461 nm, respectively. The fluorescence intensity of Hoechst 33342 increases with the pH of the solvent. The unbound dye fluoresces in the 510–540 nm range. The green fluorescence of unbound dye may be observed when an excessive dye concentration is used or the sample is insufficiently washed out. The dye has a considerable Stokes shift between the excitation and emission spectrum, making it worthwhile in multicolor labeling experiments.

The additional ethyl group in Hoechst 33342 renders it more cell-permeable than DAPI and other Hoechst dyes. Hoechst 33342 exhibits a 10-fold greater cell permeability than Hoechst 33258. Hoechst 33342 is also less toxic than DAPI, which ensures a higher viability of stained cells.

Hoechst 33342 is used extensively in fluorescence microscopy and flow cytometry for staining chromosomes and nuclei in live and fixed cells. The dye is often used to distinguish condensed pycnotic nuclei in apoptotic cells and cell sorting.

Hoechst 33342 is quenched by bromodeoxyuridine (BrdU), commonly used to detect dividing cells. When BrdU is integrated into DNA, the bromine is supposed to deform the minor groove so that Hoechst dyes cannot reach their optimal binding site. This property of Hoechst 33342 is used to study cell-cycle progression.

The commonly used dye concentration to stain bacteria or eukaryote cells is 0.1–10 μg/mL.

We offer Hoechst 33342 as a powder (1H010), a concentrated 10 mg/mL (2G010), and a ready-to-use (coming soon) aqueous solutions.

Absorption and emission spectra of Hoechst 33342 (DNA–dye complex)

Absorption and emission spectra of Hoechst 33342 (DNA–dye complex)

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General properties

Appearance: orange solid, yellow solution
Molecular weight: 561.95
CAS number: 23491-52-3
Molecular formula: C27H31Cl3N6O
IUPAC name: 2′-(4-Ethoxyphenyl)-5-(4-methyl-1-piperazinyl)-2,5′-bi-1H-benzimidazole trihydrochloride
Solubility: water, DMSO, DMF
Quality control: NMR 1H and HPLC-MS (95+%)
Storage conditions: 24 months after receival at -20°C in the dark. Transportation: at room temperature for up to 3 weeks. Desiccate.
MSDS: Download
Product specifications

Spectral properties

Excitation/absorption maximum, nm: 351 (complex)
Emission maximum, nm: 461 (complex)
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