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| Product Name | Hoechst 33258 Stain Solution (1 mg/mL) |
| Catalog No. | RESK-0539 |
| 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. It is suitable for nuclear staining of fixed cells or tissues as well as living cells or tissues. This product is supplied as a ready-to-use solution at a defined concentration of 1 mg/mL. For nuclear staining, the recommended working concentration is 0.5-10 μg/mL. A powder form of Hoechst 33258 is also available separately. |
| Intended Use | Apoptosis detection by fluorescence microscopy or flow cytometry; nuclear staining of fixed or living cells and tissues; routine nuclear staining and 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 Concentration: 1 mg/mL 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, dilute the 1 mg/mL stock solution to a recommended working concentration of 0.5-10 μg/mL. |
| Components / Formulation | Hoechst 33258 solution at 1 mg/mL. |
| Storage Conditions | Store at 4 °C, protected from light. |
| Shelf Life | 1 year for the 1 mg/mL Hoechst 33258 stock solution when stored at 4 °C protected from light. |
| Package Specifications | 1 mL |
| Product Form | Solution |
| Quality Control | Purity >=95% (HPLC). |
| Purity | >=95% (HPLC) |
| Concentration | 1 mg/mL; recommended working concentration for nuclear staining: 0.5-10 μg/mL. |
| Molecular Weight | 533.88 |
| Shipping Conditions | Shipped with wet ice. |
| Expiration Date / Stability | The 1 mg/mL Hoechst 33258 stock solution is valid for 1 year when stored at 4 °C protected from light. |
| Regulatory / Compliance | For research use only. |
| Compatibility | Detectable with DAPI filter sets or blue GFP filter sets. Hoechst 33258 and Hoechst 33342 are very similar; Hoechst 33342 is somewhat less toxic to cells, so Hoechst 33258 is generally used to stain cells after fixation, whereas Hoechst 33342 can be applied directly to living cells. |
| Application Notes / Precautions | Hoechst 33258 and Hoechst 33342 differ only slightly; Hoechst 33342 is less cytotoxic. Hoechst 33258 is generally used for staining after cell fixation, while Hoechst 33342 can stain living cells directly. 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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