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| Product Name | Hoechst 33258 |
| Catalog No. | RESK-0538 |
| Description | Hoechst 33258 is a blue fluorescent dye that can penetrate cell membranes. It releases intense blue fluorescence upon intercalation into double-stranded DNA and exhibits low toxicity to cells. Hoechst 33258 is commonly used for apoptosis detection, with stained samples observed by fluorescence microscopy or analyzed by flow cytometry; it can also be used for routine nuclear staining or general DNA staining. For nuclear staining, the recommended working concentration of Hoechst 33258 is 0.5-10 μg/mL. |
| Intended Use | Apoptosis detection by fluorescence microscopy or flow cytometry; routine nuclear staining of cells; general DNA staining. For research use only. |
| Principle / Technology | Hoechst 33258 is a cell membrane-permeable bisbenzimide dye that intercalates into double-stranded DNA and, upon binding, emits intense blue fluorescence. Fluorescence spectra: Ex/Em = 346/460 nm for free Hoechst 33258; Ex/Em = 350/460 nm for the Hoechst 33258-nucleic acid complex. The fluorescence can be excited with a xenon lamp, mercury arc lamp, or UV laser, and detected with a DAPI filter set, a blue GFP filter set, or similar blue-fluorescence filter sets. |
| Detection Method | Fluorescence microscopy or flow cytometry. Fluorescence spectra: Ex/Em = 346/460 nm for free Hoechst 33258; Ex/Em = 350/460 nm for the Hoechst 33258-nucleic acid complex. The fluorescence can be excited with a xenon lamp, mercury arc lamp, or UV laser, and detected with a DAPI filter set, a blue GFP filter set, or similar blue-fluorescence filter sets. |
| Sample Type | Living or fixed cells and tissues; cell nuclei; double-stranded DNA. |
| Performance Range / Specifications | English synonyms: 2'-(4-Hydroxyphenyl)-5-(4-methyl-1-piperazinyl)-2,5'-bi-1H-benzimidazole, trihydrochloride; bisBenzimide H 33258; HOE 33258 Molecular Formula: C25H24N6O·3HCl Molecular Weight: 533.88 CAS NO.: 23491-45-4 Fluorescence spectra: Ex/Em = 346/460 nm for free Hoechst 33258; Ex/Em = 350/460 nm for the Hoechst 33258-nucleic acid complex. The fluorescence can be excited with a xenon lamp, mercury arc lamp, or UV laser, and detected with a DAPI filter set, a blue GFP filter set, or similar blue-fluorescence filter sets. Purity: >=95% (HPLC) |
| Specificity | Binds to double-stranded DNA and emits intense blue fluorescence upon intercalation; low toxicity to cells. |
| Reaction Conditions / Protocol | For nuclear staining, the recommended working concentration of Hoechst 33258 is 0.5-10 μg/mL. The dye can be applied directly to living cells without fixation. |
| Components / Formulation | Supplied as a solid (Hoechst 33258 trihydrochloride). |
| Storage Conditions | Store at -20 °C, dry and protected from light. |
| Shelf Life | 2 years when stored at -20 °C, dry and protected from light. |
| Package Specifications | 10 mg 500 mg 1 g |
| Product Form | Solid |
| Quality Control | Purity >=95% (HPLC). |
| Purity | >=95% (HPLC) |
| Concentration | Recommended working concentration for nuclear staining: 0.5-10 μg/mL. |
| Molecular Weight | 533.88 |
| Shipping Conditions | Shipped with wet ice. |
| Expiration Date / Stability | Valid for 2 years when stored at -20 °C, dry and protected from light. |
| Regulatory / Compliance | For research use only. |
| Compatibility | Hoechst 33258 can be used together with Hoechst 33342 for nuclear staining and can be combined with lysosomal dyes for simultaneous staining of lysosomes and nuclei; the dye stains living cells directly without fixation. Detectable with DAPI filter sets or blue GFP filter sets. |
| Application Notes / Precautions | For simultaneous staining of lysosomes and nuclei, Hoechst 33258 or Hoechst 33342 is recommended for the nuclear staining step, as these dyes can stain living cells directly without fixation. Compared with DAPI (a semi-permeant to impermeant dye better suited to fixed/permeabilized cells), membrane-permeable Hoechst dyes with the same binding mechanism and fluorescence characteristics are well suited for live-cell imaging. Protect from light during storage. |
For research use only, not for clinical use.
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