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DAPI染色試薬

蛍光顕微鏡での核染色試薬(青色)

DAPI (diamidino-2-phenylindole) は二本鎖DNAの副溝(マイナーグルーブ)に選択的に結合することでDNAに未結合の状態に比べて20倍も強い蛍光を発する鮮やかな青色蛍光プローブです。このDNAに対する結合選択性と細胞透過性により、DAPIを用いた染色は細胞質由来のバックグラウンドがほとんど無い核カウンター染色法として蛍光顕微鏡では従来から用いられています。DAPIは核カウンター蛍光染色試薬として知られるHoechstに比べて、高いフォトスタビリティをもちます。最大励起波長は345nm、最大発光波長は455nmになります。DAPIストック溶液は-20℃での遮光保管で安定です。

DAPIの特性
Alternative names 4,6'-diamidino-2-phenylindole, dihydrochloride
4',6-diamidine-2-phenyl indole
Molecular mass 350.25
Excitation wavelength 345 nm (near 360 nm when bound to dsDNA)
Emission wavelength 455 nm (456-460 nm when bound to dsDNA)
Extinction coefficient ~30,000/M cm at 347 nm in methanol
CAS # 28718-90-3
Purity by HPLC >95% (most lots >98%)
Solubility Soluble in DMF, water and various non-phosphate aqueous buffers
Storage Room temperature (RT), protected from light
Reactive groups Non-covalent; binds to minor groove of double-stranded DNA

アプリケーション

  • Assaying DNA in solution (ref.4)
  • Diagnosing mycoplasmal infection of cell cultures (ref.5)
  • Measuring nuclear content and sorting chromosomes in flow cytometry (ref.6)
  • Assessing apoptosis (ref.7)
  • Detecting nuclei and organellar DNA in immunofluorescent and in situ hybridization procedures (ref.2,8)
  • Replacing ethidium bromide for staining DNA in agarose gels (ref.5,9)
  • Counterstaining nuclei in histochemical methods when red-fluorescent antibodies have been used to detect specific targets (ref.8)
  • Reports also indicate that DAPI will bind to polyphosphates and other polyanions (ref.10), dextran sulfate (ref.11) and SDS (ref.12).

参考文献

  1. Morikawa, K. and Yanagida, J. (1981). Visualization of individual DNA molecules in solution by light microscopy: DAPI staining method. J. Biochem. (Tokyo) 89:693-6.
  2. 2. Lawrence, M.E. and Possingham, J.V. (1986). Direct measurement of femtogram amounts of DNA in cells and chloroplasts by quantitative microspectrofluorometry. J. Histochem. Cytochem. 34:761-8.
  3. Kubista, M., et al. (1987). Characterization of interaction between DNA and 4', 6-diamidino-2-phenylindole by optical spectroscopy. Biochemistry 26:4545-53.
  4. Brunk, C.F., et al. (1979). Assay for nanogram quantities of DNA in cellular homogenates. Anal. Biochem. 92:497-500.
  5. Russell, W.C., et al. (1975). A simple cytochemical technique for demonstration of DNA in cells infected with mycoplasmas and viruses. Nature 253:461-2.
  6. Hammarton, T.C., et al. (2003). Stage-specific differences in cell cycle control in Trypanosoma brucei revealed by RNA interference of a mitotic cyclin. J. Biol. Chem. 278(25):22877-86.
  7. Lai, J., et al. (2003). Loss of HSulf-1 up-regulates heparin-binding growth factor signaling in cancer. J. Biol. Chem. 278(25):23107-17.
  8. Soto, P., et al. (2003). SMAD2 and SMAD7 Involvement in the post-translational regulation of Muc4 via the transforming growth factor-b and interferon-g pathways in rat mammary epithelial cells. J. Biol. Chem. 278(22):20338-44.
  9. Nairn, R.S., et al. (1982). Comparison of ethidium bromide and 4', 6'-diamidino-2-phenylindole as quantitative fluorescent stains for DNA in agarose gels. J. Biochem. Biophys. Meth. 6:95-103.
  10. Tijssen, J.P.F., et al. (1982). Localization of polyphosphates in Saccharomyces fragilis, as revealed by 4', 6-diamidino-2-phenylindole fluorescence. Biochem. Biophys. Acta 721:394-8.
  11. Allan, R.A. and Miller, J.J. (1980). Influence of S-adenosylmethionine on 4', 6-diamidino-2-phenylindole dihydrochloride (DAPI)-induced fluorescence of polyphosphate in the yeast vacuole. Can. J. Micro. 26:912-20.
  12. Kapuscinski, J. and Skoczylas, B. (1978). Fluorescent complexes of DNA with DAPI (4',6-diamidine-2-phenyl indole dihydrochloride) or DCI (4',6- dicarboxyamide-2-phenyl indole). Nucl. Acids Res. 5:3775-99.

Thermo Scientific Pierce Fluorescent Dyes are available in bulk quantities and prices.
*Trademark, see Trademark Index

製品情報
Cat # 製　品　名 容 量 参考価格
46190 4',6-Diamidino-2-phenylindole, dihydrochloride (DAPI) 10 mg 販売終了