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| Product Name | DAPI (4',6-Diamidino-2-phenylindole Dihydrochloride) |
| Catalog No. | RESK-0536 |
| Description | DAPI, also known as DAPI dihydrochloride, is a commonly used nucleic acid dye that binds to the minor groove of AT-rich sequences in double-stranded DNA, producing blue fluorescence more than 20 times stronger than that of the free dye. Compared with ethidium bromide (EB), DAPI is many times more sensitive for double-stranded DNA staining. DAPI can also stain RNA by selectively intercalating into AU sequences and emitting fluorescence. Compared with the DAPI-dsDNA complex (Ex/Em = 358 nm/461 nm), the DAPI-RNA complex has a longer maximum emission wavelength (500 nm) and a fluorescence brightness of only 20% of that of DAPI-dsDNA. Although DAPI cannot pass through the membranes of living cells, it can be driven into live cells by increasing its concentration. DAPI has a very high level of photobleaching resistance and can be used to detect yeast mitochondrial DNA, chloroplast DNA, viral DNA, mycoplasma DNA, and chromosomal DNA. The characteristic blue fluorescence of DAPI makes it a very common partner for green, yellow, or red fluorescent dyes in multicolor fluorescence labeling techniques in cell biology. It is therefore also frequently used as a nuclear and chromosomal counterstain in apoptosis detection, RNA in situ hybridization, and direct or indirect immunodetection. Stained samples can be observed by fluorescence microscopy or analyzed by flow cytometry. The recommended working concentration is 0.5-10 μg/mL. This product is supplied as a powder. |
| Intended Use | Fluorescent staining of double-stranded DNA; nuclear and chromosomal counterstaining in apoptosis detection, RNA in situ hybridization, and direct or indirect immunodetection; detection of yeast mitochondrial DNA, chloroplast DNA, viral DNA, mycoplasma DNA, and chromosomal DNA; multicolor fluorescence labeling together with green, yellow, or red fluorescent dyes. For research use only. |
| Principle / Technology | DAPI binds to the minor groove of AT-rich sequences in double-stranded DNA, yielding a DAPI-DNA complex with blue fluorescence more than 20 times stronger than that of free DAPI (Ex/Em = 358 nm/461 nm). DAPI also stains RNA by selectively intercalating into AU sequences; the DAPI-RNA complex has a longer maximum emission wavelength (500 nm) and only 20% of the brightness of the DAPI-dsDNA complex. |
| Detection Method | Fluorescence microscopy or flow cytometry. Fluorescence spectra: Ex/Em = 340 nm/488 nm for free DAPI; Ex/Em = 358 nm/461 nm for the DAPI-DNA complex. |
| Sample Type | Fixed and permeabilized cells and tissues (bright nuclear labeling); double-stranded DNA; RNA; yeast mitochondrial DNA; chloroplast DNA; viral DNA; mycoplasma DNA; chromosomal DNA. DAPI is considered a semi-permeant to impermeant dye, so staining of living cells is inconsistent and is not recommended; Hoechst dyes are suggested for live-cell staining. |
| Performance Range / Specifications | English synonyms: 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride; DAPI dihydrochloride; 4',6-Diamidino-2-phenylindole, dihydrochloride Molecular Formula: C16H15N5·2HCl Molecular Weight: 350.25 CAS NO.: 28718-90-3 Purity (HPLC): >=95% Fluorescence spectra: Ex/Em = 340 nm/488 nm for DAPI; Ex/Em = 358 nm/461 nm for DAPI-DNA |
| Sensitivity / LOD | Significantly more sensitive than ethidium bromide (EB) for double-stranded DNA staining; upon binding to dsDNA, DAPI produces blue fluorescence more than 20 times stronger than that of the free dye. |
| Specificity | Binds preferentially to AT-rich sequences in the minor groove of double-stranded DNA; also selectively intercalates into AU sequences of RNA. |
| Reaction Conditions / Protocol | Recommended working concentration: 0.5-10 μg/mL. For staining, the culture medium may be removed first and 100 μL of staining solution added directly. This product is not recommended for staining living cells; for live-cell staining, Hoechst dyes are recommended. |
| Components / Formulation | Supplied as a powder. |
| Storage Conditions | Store the powder at 4 °C, protected from light. |
| Shelf Life | 3 years when stored at 4 °C protected from light. |
| Package Specifications | 10 mg |
| Product Form | Powder |
| Quality Control | Purity >=95% (HPLC). |
| Purity | >=95% (HPLC) |
| Concentration | Recommended working concentration: 0.5-10 μg/mL. |
| Molecular Weight | 350.25 |
| Shipping Conditions | Shipped at room temperature. |
| Expiration Date / Stability | Valid for 3 years when the powder is stored at 4 °C protected from light. |
| Regulatory / Compliance | For research use only. |
| Compatibility | Commonly combined with green, yellow, or red fluorescent dyes for multicolor fluorescence labeling in cell biology; suitable as a nuclear and chromosomal counterstain in apoptosis detection, RNA in situ hybridization, and direct or indirect immunodetection. Hoechst 33342 or Hoechst 33258, which share the same wavelengths and nucleic acid binding mode but show very good cell membrane permeability, can serve as alternatives for live-cell nuclear labeling. |
| Application Notes / Precautions | DAPI is regarded as a semi-permeant/impermeant nuclear dye; nuclear staining performance depends on the cell line. Some cell lines can be stained with DAPI, some cannot, and some give inconsistent labeling. For live-cell staining, Hoechst 33342 or Hoechst 33258 are recommended alternatives with the same wavelengths and nucleic acid binding mode but excellent membrane permeability. DAPI has a very high level of photobleaching resistance. Protect from light during storage. |
For research use only, not for clinical use.
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