The Thermo Scientific DRAQ5* Fluorescent Probe is a far-red DNA stain for live or fixed cells and easy multiplexing with other fluorophores, such as cells expressing green fluorescent protein (GFP) fusion proteins.
DRAQ5* Fluorescent Probe is a far-red DNA stain for fluorescent cellular imaging applications with live or fixed cells. Because of its far-red excitation and emission, the DRAQ5 Stain can be multiplexed with many other fluorophores. It is commonly chosen to counterstain nuclei and measure DNA content in cell proliferation studies involving cells expressing green fluorescent protein (GFP) fusion proteins. DRAQ5 Fluorescent Probe is compatible with many existing protocols across a wide range of instrumentation platforms.
Highlights:
- DRAQ5 dye – a red-to-infrared fluorescent stain specific for DNA (nuclei and DNA quantitation)
- Convenient – provided as easy-to-use DRAQ5 dye solutions (5mM)
- Cellular imaging – lipophilic and membrane permeable dye is effective for live-cell and fixed-cell staining and quantitation of DNA content
- Counterstain – ideal for use alongside other fluorophore reporters for fluorescence microscopy or high-content screening (HCS)
Applications:
- Flow cytometry
- Flow and laser scanning cytometry
- Live cell imaging
- Cell based assays
Properties of DAPI Fluorescent Dye:
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Structure
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Proprietary
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Alternative names
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Anthraquinone compound
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Chemical name(s)
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1,5-bis{[2-(di-methylamino) ethyl]amino}-4, 8-dihydroxyanthracene-9,10-dione
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Molecular formula
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Not available
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Molecular weight
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412.54
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Excitation wavelength
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647nm line optimal (Exλmax = 646nm)
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Emission wavelength
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> 665nm to infra-red > 800nm
(Emλmax = 681nm / 697nm intercalated with dsDNA)
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Extinction coefficient
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Not available
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CAS #
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Not available
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Purity
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> 99%
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Solubility
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Soluble at working concentrations in water and typical buffers
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Storage
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Store at 4°C or room temperature, protected from light
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Reactive groups
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None; binds DNA A-T minor groove, mimicking histone 2b distribution, resulting in 16nm red-shift
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Product Details:
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| DRAQ5* Dye is a red fluorescent DNA stain. HepG2 cells stained with Thermo Scientific DRAQ5 Fluorescent Probe. |
Fluorescence cell-based assays, such as flow cytometry, in-cell ELISA, fluorescence microscopy and high-content imaging require a fluorescent label to identify individual cells. When using multiple fluorescent probes to detect different cellular targets or activities, each probe must have a fluorescent spectrum different than the other probes. The blue-fluorescent DNA-binding probes, Hoechst and DAPI are frequently used; however, these probes cannot be used when UV illumination is unavailable or other blue-emitting fluorescent probes are used. Therefore, nuclear probes that emit in a color other than blue are useful for cell identification and counting, and for determining nuclear morphology and DNA content.
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| DRAQ5 Dye provides DNA counterstaining in experiments involving blue fluorescent probes. Glutathione (GSH) is rapidly depleted in A549 cells upon 300μM menadione treatment for 30 minutes at 37ºC, resulting in decreased staining by blue-fluorescent monochlorobimane (MBCL) and monobromobimane (MBBR). Fluorescence intensity in separate blue and red channels allows quantitation. Cell images were acquired on the Thermo Scientific Cellomics ArrayScan HCS Reader with a 20X objective lens. |
DRAQ5 Fluorescent Probe emits in the far-red region, is lipophilic and crosses cell and nuclear membranes in live and fixed cells and tissues for rapid DNA staining. This stain is water-soluble, supplied ready to use and does not require RNase, cell lysis, or a washing step, making it compatible with automation. Because DNA staining is stoichiometric, the DRAQ5 Fluorescent Probe can be used for DNA content analysis in cell proliferation studies. DRAQ5 Fluorescent Probe is usually added as the last stain in a labeling procedure because no washing is required. Alternatively, add this probe in assay medium for a live cell assay.
References:
- Edward, R. (2009). Use of DNA-specific anthraquinone dyes to directly reveal cytoplasmic and nuclear boundaries in live and fixed cells. Mol Cells 27:391-6.
- Martin, R.M., et al. (2005). DNA labeling in living cells. Cytometry 67A:45-52.
- Smith, P.J., et al. (1999). A novel cell permeant and far red-fluorescing DNA probe, DRAQ5, for blood cell discrimination by flow cytometry. J Immunol Methods 229(1-2):131-9.
- Smith, P.J., et al. (2000). Characteristics of a novel deep red/infrared fluorescent cell-permeant DNA probe, DRAQ5, in intact human cells analyzed by flow cytometry, confocal and multiphoton microscopy. Cytometry 40(4):280-91.
- Swerts, K., et al. (2007). DRAQ5: Improved flow cytometric DNA content analysis and minimal residual disease detection in childhood malignancies. Clin Chim Acta 379:154-157.
- Wiltshire, M., et al. (2000). A novel deep red/low infrared fluorescent flow cytometric probe, DRAQ5NO, for the discrimination of intact nucleated cells in apoptotic cell populations. Cytometry 39(3):217-23.
- Yuan, C.M., et al. (2004). DRAQ5-based DNA content analysis of hematolymphoid cell subpopulations discriminated by surface antigens and light scatter properties. Cytometry 58:4752.
Related Products:
DyLight Reactive Fluors
All Fluorescent Labeling Reagents – including AMCA, FITC, and TRITC
Immunohistochemistry Reagents – including DAPI and Hoechst dyes
Cellomics High-Content Screening (HCS) Reagents
Related Resources:
Review of Immunohistochemistry (IHC) Methods
Review of IHC Counterstain Methods
Tech Tip #31 – Calculate dye:protein (F/P) molar ratios
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