Hoechst 33258 is a fluorescent dye based on Hoechst 33258 [4-(5-(4-methylpiperazin-1-yl )-1H,1’H-[2,5′-biphenyl[D]imidazolyl]-2′-yl )phenol], which is widely used in cellular imaging for the staining of DNA and cell nuclei. The detection principle is that Hoechst 33258 penetrates the cell membrane and binds to the adenine-thymine (A-T) groove of dsDNA to stain the nucleus. Hoechst 33258 has a low fluorescence background and emits an intense blue fluorescence (maximum excitation and emission of Ex = 352 nm and Em = 461 nm, respectively) when bound to dsDNA. Hoechst 33258 staining is commonly used for apoptosis detection, general nuclear staining or routine DNA staining, and can be visualized by fluorescence microscopy or flow cytometry. Hoechst 33258 is easy to use, simply dilute Hoechst 33258 solution to the appropriate concentration, cover the sample and incubate for a certain period of time to observe the bright blue fluorescence. This product can be widely used for nuclear staining of living cells, fixed cells or tissues, paraffin sections, apoptosis and cell cycle analysis; due to the poor permeability of Hoechst 33258 to the cell membrane of living cells, the intensity of the blue fluorescence is weak when staining healthy living cells, but for apoptotic cells or cells with damaged cell membranes, it will emit bright blue fluorescence; if you need bright blue fluorescence when staining healthy living cells, you can use Hoechst 33342 Solution (Code No. orb3229983). For apoptotic cells or cells with damaged cell membranes, bright blue fluorescence will be emitted; if bright blue fluorescence is required for staining healthy living cells, Hoechst 33342 Solution (Code No. orb3229983) can be used.
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Application Notes
1. This product is a ready-to-use dye, easy to operate, when staining fixed cells or paraffin section samples, it can be used to dilute it to the working concentration (0.5 ~ 10 μg / ml) with PBS or cell culture solution, add it to the cells, incubate for a certain period of time at room temperature and then observe; when staining live cells, the solution can be added directly to the cell culture solution according to a certain ratio, without the need to collect the cells or wash the cells, and then incubate for a certain period of time at 37℃ to observe. When staining live cells, the solution can be added directly to the cell culture medium in a certain proportion, without collecting cells or washing them, and then incubated at 37℃ for a certain time for observation. 2. This product can penetrate the cell membrane, low toxicity to cells, suitable for live cells, fixed cells or tissues, paraffin sections and other samples of cell nuclear staining experiments and apoptosis detection. 3. Phenol red and serum have no effect on Hoechst 33258 staining.
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