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| Product Name | Hoechst 33342 Stain Solution (1 mg/mL) |
| Catalog No. | RESK-0541 |
| Description | Hoechst 33342 is a cell membrane-permeable blue fluorescent dye that emits strong blue fluorescence upon binding to double-stranded DNA and shows low cytotoxicity to cells. It is commonly used for apoptosis detection, with stained cells examined by fluorescence microscopy or analyzed by flow cytometry, and is also suitable for routine nuclear counterstaining and general DNA staining. 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 format of Hoechst 33342 is also available as a separate product. |
| Intended Use | Qualitative and quantitative fluorescence staining of nuclear DNA in live or fixed cells; apoptosis detection; routine nuclear counterstaining and general DNA staining, analyzed by fluorescence microscopy or flow cytometry. |
| Principle / Technology | Hoechst 33342 is a bisbenzimide dye that crosses intact cell membranes and binds to double-stranded DNA. Upon DNA binding its blue fluorescence is strongly enhanced. Unlike DAPI, which is semi-permeant to impermeant and gives inconsistent labeling of live cells, Hoechst 33342 is fully membrane-permeant and is therefore well suited for live-cell imaging, while performing like DAPI on fixed samples. |
| Detection Method | Fluorescence microscopy; flow cytometry |
| Sample Type | Live cells (e.g., A549); fixed and permeabilized cells or tissues |
| Performance Range / Specifications | English synonym: 2'-(4-Ethoxyphenyl)-5-(4-methyl-1-piperazinyl)-2,5'-bi-1H-benzimidazole, trihydrochloride Molecular formula: C27H28N6O·3HCl Molecular weight: 561.93 CAS No.: 23491-52-3 Fluorescence spectra: Ex/Em = 346/460 nm for free Hoechst 33342; Ex/Em = 350/461 nm for the Hoechst 33342-nucleic acid complex Purity: ≥95% (HPLC) |
| Specificity | Selectively binds double-stranded DNA with low cytotoxicity; suitable for live-cell nuclear labeling. |
| Reaction Conditions / Protocol | Dilute the 1 mg/mL stock to a working concentration of 0.5-10 μg/mL and incubate cells before analysis by fluorescence microscopy or flow cytometry. Live cells can be stained directly without fixation. No fixed reference values are available for specific cell lines such as A549; a pilot experiment within the recommended concentration range is advised to determine the optimal concentration and staining time. |
| Components / Formulation | Hoechst 33342 stock solution, 1 mg/mL; ≥95% purity (HPLC) |
| Storage Conditions | Store at 4°C, protected from light. |
| Shelf Life | 1 year when stored at 4°C, protected from light (1 mg/mL stock solution). |
| Package Specifications | 1 mL |
| Product Form | Solution (1 mg/mL) |
| Purity | ≥95% (HPLC) |
| Concentration | 1 mg/mL |
| Molecular Weight | 561.93 |
| Shipping Conditions | Shipped on wet ice. |
| Expiration Date / Stability | The 1 mg/mL stock solution is stable for 1 year at 4°C, protected from light. |
| Regulatory / Compliance | For research use only. |
| Compatibility | Fluorescence can be excited with a xenon lamp, mercury arc lamp, or UV laser, and detected using a DAPI filter set or a blue GFP filter set. |
| Application Notes / Precautions | Recommended working concentration for nuclear staining: 0.5-10 μg/mL. Protect from light during storage and handling. For specific cell lines without reference data, run a pilot experiment within the recommended concentration range. |
| Recognition Site | Double-stranded DNA (nuclear DNA) |
For research use only, not for clinical use.
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