PureBlu™ Hoechst 33342 Nuclear Staining Dye #1351304

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Molecular formula C27H28N6O·3HCl
Molecular weight 561.93
Excitation/emission wavelengths 350/461 nm
CAS Number 23491-52-3
Purity >95% (HPLC purified)
Solubility Soluble in water and dimethyl sulfoxide (DMSO)
Storage and use –20ºC
Temperature stability Stable for 2 years at –20°C
After resuspension: stable for 1 year at –20°C or 6 months at 2–8°C

Use PureBlu Hoechst 33342 Dye for routine nuclear staining in fluorescence microscopy and cell imaging applications. PureBlu Hoechst 33342 Dye is a highly pure formulation of Hoechst 33342 in a user-friendly format allowing easy preparation of the working solution, with no weighing step and only a single dilution after resuspension.

Hoechst 33342 is a well-characterized blue-emitting fluorescent compound widely utilized for nuclear staining. Hoechst 33342 permeates cell membranes and binds to the minor groove of A/T-rich dsDNA sequences, thus preferentially staining nuclei (Figure 1).

Hoechst 33342 binding dsDNA

Fig. 1. Hoechst 33342 binding dsDNA.
 

When bound to dsDNA, Hoechst 33342 has a maximum excitation wavelength in the ultraviolet range (350 nm), and the dye can be optimally detected in the blue channel with an emission maximum of 461 nm (Figure 2). The characteristic Stokes shift between excitation and emission wavelengths is fairly wide for Hoechst 33342, making this dye an optimal choice when good spectral separation is desired to reduce fluorescence interference, for example, in chromatin counterstaining for immunofluorescence microscopy.


Hoechst 33342 emission spectrum

Fig. 2. Hoechst 33342 excitation and emission spectra.
 

PureBlu Hoechst 33342 Nuclear Staining Dye is compatible with fixed and unfixed cells. Compared to DAPI, it exhibits greater permeability toward intact cell membranes and is preferred when conducting experiments with unfixed samples.

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Waste Electrical and Electronic Equipment ("WEEE Directive")

This product contains electrical or electronic material. Learn more about Bio-Rad's EU Recycle Program