D9542
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D9542
Sigma
4′,6-Diamidino-2-phenylindole dihydrochloride
powder, ≥98% (HPLC and TLC)
| Synonym: | 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride, DAPI dihydrochloride |
| CAS Number: | 28718-90-3 |
| Linear Formula: | C16H15N5 · 2HCl |
| Molecular Weight: | 350.25 |
| Beilstein Registry Number: | 4894417 |
| EC Number: | 249-186-7 |
| MDL number: | MFCD00012681 |
| PubChem Substance ID: | 24894305 |
Description
| Frequently Asked Questions | Live Chat and Frequently Asked Questions are available for this Product. |
| Biochem/physiol Actions | Cell permeable fluorescent minor groove-binding probe for DNA. Binds to the minor groove of double-stranded DNA (preferentially to AT rich DNA), forming a stable complex which fluoresces approximately 20 times greater than DAPI alone. |
| Caution | Protect from light. |
| Application | DAPI is several times more sensitive than ethidium bromide for staining DNA in agarose gels. It may be used for photofootprinting of DNA, to detect annealed probes in blotting applications by specifically visualizing the double-stranded complex, and to study the changes in DNA and analyze DNA content during apoptosis using flow cytometry. DAPI staining has also been shown to be a sensitive and specific detection method for mycoplasma. |
Properties
| assay | ≥98% (HPLC and TLC) |
| form | powder |
| fluorescence | λex 340 nm; λem 488 nm (nur DAPI) |
| λex 364 nm; λem 454 nm (DAPI-DNA-Komplex) | |
| solubility | H2O: 20 mg/mL heat or sonication may be required. Solutions stored in the dark at room temperature or 4 °C should be stable for 2 to 3 weeks. |
| PBS: insoluble | |
| abs. | ε |
| storage temp. | 2-8°C |
Safety
| Personal Protective Equipment | Eyeshields, Gloves, type N95 (US), type P1 (EN143) respirator filter |
| Hazard Codes | Xi |
| Risk Statements | 36/37/38 |
| Safety Statements | 26-36 |
| WGK Germany | 3 |
References
| reference | Naimski, P., Quantitative fluorescent analysis of different conformational forms of DNA bound to the dye, 4', 6-diamidine-2-phenylindole, and separated by gel electrophoresis. Anal. Biochem. 106, 471, (1980) |
| Otto, F.J., High-resolution analysis of nuclear DNA employing the fluorochrome DAPI. Methods Cell Biol. 41, 211-217, (1994) | |
| Kapuscinski, J., DAPI: a DNA-specific fluorescent probe. Biotech. Histochem. 70, 220-233, (1995) | |
| Darzynkiewicz, Z. and Li, X., Cotter, T.G. and Martin, S.J., ed., Measurements of cell death by flow cytometry, in Techniques in Apoptosis: A User's Guide. Cytometry London, U.K. , (1996), 91-92 | |
| Uphoff, C.C., et al., Mycoplasma contamination in human leukemia cell lines. I. Comparison of various detection methods. J. Immunol. Methods 149, 43-53, (1992) | |
| Jeppesen, C.,, Nielsen, P.E, Photofootprinting of drug-binding sites on DNA using diazo- and azido-9-aminoacridine derivatives. Eur. J. Biochem. 182, 437, (1989) | |
| Zaitsev, E.N., Kowalczykowski, S.C., Binding of double-stranded DNA by Escherichia coli RecA protein monitored by a fluorescent dye displacement assay. Nucleic Acids Res. 26, 650-654, (1998) | |
| Collins, J.A., Major DNA fragmentation is a late event in apoptosis. J. Histochem. Cytochem. 45, 923, (1997) | |
| Hotz, M.A., et al., Flow cytometric detection of apoptosis: comparison of the assays of in situ DNA degradation and chromatin changes. Cytometry 15, 237-244, (1994) | |
| Uphoff, C.C., et al., Sensitivity and specificity of five different mycoplasma detection assays. Leukemia 6, 335-341, (1992) |
