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| Product Name | DAPI Nuclear Counterstaining Solution for Fluorescence Microscopy |
| Catalog No. | CCAT-HMM-0051 |
| Description | A ready-to-use 4',6-diamidino-2-phenylindole (DAPI) solution for the fluorescent counterstaining of nuclei in fixed-cell and tissue section preparations. DAPI forms a highly fluorescent complex with double-stranded DNA that exhibits a strong blue emission, providing a universal nuclear marker for multicolor immunofluorescence experiments. |
| Intended Use | Nuclear counterstaining in fixed-cell fluorescence microscopy, immunohistochemistry, TUNEL apoptosis detection, and DNA content-based cell cycle analysis for biomedical research applications. |
| Principle / Technology | DAPI (Ex/Em: 358/461 nm when bound to DNA) intercalates and binds selectively to A-T rich regions in the minor groove of double-stranded DNA; the fluorescence quantum yield increases approximately 20-fold upon binding; the dye requires cell fixation and permeabilization to penetrate intact plasma membranes, unlike the cell-permeable Hoechst dyes. |
| Detection Method | Fluorescence microscopy with DAPI/UV or blue filter set (Ex 330–380 nm, Em 420–470 nm); also compatible with violet (405 nm) laser excitation in confocal microscopy |
| Sample Type | Fixed and permeabilized adherent cells on coverslips or chamber slides, formalin-fixed paraffin-embedded tissue sections, frozen tissue sections, chromosome spreads, isolated nuclei |
| Performance Range / Specifications | Staining concentration: 0.1–1 μg/mL; staining time: 5–15 minutes at room temperature; fluorescence excitation maximum 358 nm, emission maximum 461 nm; nuclear staining persists for months when mounted in antifade medium |
| Sensitivity / LOD | Detection of individual nuclei at DAPI concentration as low as 0.05 μg/mL with sufficient fluorescence for high-magnification oil immersion objectives |
| Specificity | DAPI binds preferentially to consecutive A-T base pairs in double-stranded DNA; minimal binding to RNA (fluorescence emission shifts to approximately 500 nm with RNA); negligible mitochondrial DNA staining at standard concentrations |
| Reaction Conditions / Protocol | For fixed-cell preparations: apply 300 nM DAPI in PBS or mounting medium; incubate 5–15 minutes at room temperature protected from light; wash briefly with PBS; mount in antifade medium. For tissue sections: apply DAPI-containing mounting medium directly; incubate briefly; coverslip and seal; optimal counterstaining is achieved by including DAPI in the final mounting medium for convenience |
| Components / Formulation | DAPI dihydrochloride solution (300 nM or 1 μg/mL) in PBS or antifade mounting medium, supplied ready-to-use in dropper bottle; also available as 14 mM (5 mg/mL) stock solution for custom dilution in mounting media |
| Storage Conditions | Aqueous stock solution: 2–8°C protected from light, stable 12 months; lyophilized solid: -20°C protected from light and moisture, stable 36 months |
| Shelf Life | 36 months from date of manufacture for lyophilized form |
| Package Specifications | 10 mL (300 μM ready-to-use), 2 mL (10 mg/mL stock), 10 mg or 100 mg lyophilized |
| Product Form | Ready-to-use aqueous solution or concentrated stock requiring dilution |
| Quality Control | Each lot tested for nuclear staining specificity on HeLa cell preparations; DNA staining intensity verified; endotoxin tested for live-cell compatible formulations (<0.1 EU/mL) |
| Key Features | Gold-standard nuclear counterstain for multicolor fluorescence imaging; broad UV excitation compatible with mercury arc, xenon, and LED light sources; high photostability when mounted in antifade media; intense blue fluorescence distinct from common green/red fluorophores enabling multiplexing |
For research use only, not for clinical use.
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