DRAQ7™ Far-Red Fluorescent DNA Dye is a membrane impermeant dye for the rapid and convenient staining of the nuclear DNA of cells, tissues, and organisms including dead, permeabilized or fixed mammalian cells. It is ideal as a dead/live gate to assess sample quality, in apoptosis and cytotoxicity studies, and for immunofluorescence, immunohistochemistry, and high content screening applications.
- DRAQ7 differentially stains both the nucleus and cytoplasm in dead, permeabilized or fixed mammalian cells, making it ideal for a HCS translocation (or Redistribution™ / Transfluor™) assay. The intensity thresholding for the two compartments improves an in silico “dilation” while providing morphometrics.
- DRAQ7 directly and stoichiometrically reports DNA content in cells that are dead, permeabilized or fixed. Because DRAQ7 is a far-red emitting DNA dye, this can be easily multiplexed with other parameters, like cell surface phenotype.
- DRAQ7 is supplied as an aqueous, ready-to-use solution. The 1 ml vial (cat. no. DR71000) is sufficient for 200 flow cytometry tests, 200 to 300 slides, or 6,000 microtiter plate wells.
Identification of Dead Cells / Membrane-Compromised Cells:
Live Cell Membrane-Impermeant
The DRAQ7™ reagent only stains nuclei in dead and permeabilized cells. DRAQ7 does not enter intact, live cells. Cells may be fixed or permeabilized for use of DRAQ7 as a nuclear counterstain.
Far-red emission
The far-red emission (665 nm and beyond) of DRAQ7 permits new combinations of dyes to be used in the understanding of cell death and apoptosis. DRAQ7 is ideal for use with GFP and all visible range-fluor tagged ligands, and there is no emission overlap with PE and homologues.
Visible Range Excitation
DRAQ7 can be excited by the blue, green or red light in a flow cytometer and red light in imaging instruments, but no UV excitation is required.
Non-toxic in long-term culture
With negligible toxicity, Draq7 can be used in RNAi knock-down and long-term, dynamic viability assays to report cytotoxicity and apoptosis.
Nucl:cyto Visualization
DRAQ7 demonstrates a nucl:cyto staining differential which is useful for translocation tracking and cell morphometrics in in-vitro toxicology studies.
Nuclear Counterstaining with Fixed Cells & Tissue Sections:
For HCS and microscopy, DRAQ7™ is an ideal far-red DNA counterstain for fixed cell and tissue preparations in IF/IHC, high content screening, and cell-based assays.
- Simplify HCS assay development: far-red labeling nuclei of fixed cells offers the perfect HCS counterstain; dual Nucl:Cyto differential labeling is ideal for translocations.
- Make compound screens 10% more effective: Visible range excitation avoids UV "white-out" from coincidental natural UV fluorescence of ca. 10% of compounds in a library when using DAPI / Hoechst dyes.
- Double the speed of a screening assay: far-red emission is ideal for GFP and YFP ligands - spectral separation allows simultaneous acquisition of signals. A typical 12 week GPCR screen can be reduced to 6 weeks!!
- Spectrally compatible with UV excited fluors
- Obviates the need for compensation when used with FITC/GFP and PE tagged probes
Chemical Features:
- membrane-impermeant
- Anthraquinone compound
- Dark blue crystalline solid / solution (as supplied)
- pure synthetic compound with high affinity for DNA
- stable at room temperature
- stable under normal lighting conditions- very low photobleaching
- water soluble - compatible with buffers / media in biological pH range
- provided in a convenient ready-to-use formulation
Buy DRAQ7: Click “Add to Cart” above to purchase DRAQ7™ (0.3mM, 1 mL), cat. # DR71000, or call ICT at 800-829-3194.
DRAQ7™ is a trademark of Biostatus Ltd. DRAQ7 is intended for research purposes only.
Sample Protocol:
DRAQ7™ - EXAMPLE PROTOCOLS FOR FLOW CYTOMETRY, MICROSCOPY AND HCS TECHNOLOGIES - http://www.biostatus.com/files/technical_documents/DRAQ7%20Example%20Protocols%2006June2010.pdf
Typical use:
Flow Cytometry
200 assays - 5ul of DRAQ7 in 500ul (end concentration 3uM)
Imaging
300 assays - 3.33ul of DRAQ7 in 200ul (end concentration 5uM)
Table 1: Ready reckoner for volumes of DRAQ7™ (0.3mM) required for various cell concentrations:
|
Cell sample preparation: |
Volume of DRAQ7™ (as supplied) required for a concentration of: |
|||
|
No. of cells: |
in volume: |
3 µM |
5 µM |
10 µM |
|
5 x 105 |
1000 µl |
10 µl |
17 µl |
34 µl |
|
2.5 x 105 |
500 µl |
5 µl |
8.5 µl |
17 µl |
|
1 x 105 |
200 µl |
2 µl |
3.4 µl |
6.8 µl |
|
5 x 104 |
100 µl |
1 µl |
1.7 µl |
3.4 µl |
See Biostatus's website for more details: http://www.biostatus.com/product/draq7_new/example_protocols/
DRAQ7™ is a trademark of Biostatus Ltd. DRAQ7 is intended for research purposes only.
Multiplex for cell death with FLICA© and DRAQ7™
Jurkat cells were treated with either a placebo or staurosporine for 4 hours. After treatment, FLICA© FAM-VAD-FMK poly caspase reagent and DRAQ7™ vital dye were added to the culture media to label apoptotic and necrotic cells. Dot plots of negative and positive control samples show a clear shift from non-apoptotic cells (Q2-LL) to cells bearing either active caspases (Q2-LR), compromised cell membranes (Q2-UL), or both (Q2-UR).
Intensity-threshold segmentation of formaldehyde-fixed U2OS cells stained with DRAQ7. Note: when used with fixed cells, DRAQ7 functions as a fluorescent dsDNA stain. To use DRAQ7 as a vital dye, cell samples must remain unfixed.
Camera image from an epifluorescence microscope was segmented to view dsDNA fluorescence, nuclear region, and cytoplasm. Left panel: false-coloured grey-scale image. Centre panel: intensity-threshold segmentation to derive nuclear region. Right panel: intensity-threshold segmentation of remaining regions (for cytoplasm), excluding nuclear area derived in centre panel using a morphometric filter, and then re-combined to provide both nuclear and cytoplasmic segments.
| Excitation / Emission: | 633 & 647 nm line optimal / >665 nm to infra-red >800 nm
The graphs below show standard absorbance and fluorescence emission spectra for DRAQ7™ in aqueous solution for a range of different excitation wavelengths. As can be seen, DRAQ7™ can be effectively excited by 488nm systems. In these circumstances, we recommend that a long band pass filter be used - to capture as many emitted photons as possible. Excitation: 633 & 647 nm line optimal (Exλmax 599/644 nm) ; 488, 514 and 568 lines, sub-optimal (only by flow cytometry) Emission: >665 nm to infra-red >800 nm (Emλmax 678 nm / 694 nm intercalated with dsDNA)
Multi-wavelength imaging with UV / vis fluorochromes
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| Method of Analysis: | Flow Cytometer, Fluorescence Microscope |
| Types of Samples: | cell culture |
| Storage: | 2°-8° C, Ships at Ambient Temperature, Ships Overnight (Domestic), 2-Day, 3-Day Shipping Available for orders placed by phone, fax, or email, Do Not Freeze |
| MSDS: |
Can I use DRAQ5™ or DRAQ7™ with anti-fade mountants such as ProLong Gold, Fluoroshield, Polymount Aqua or Vectashield?
Yes. These DNA-binding anthraquinone dyes are perfectly compatible with anti-fade mountants. Generally, the cells are counterstained before the application of the mountant.
However, Serda, et al (2009) doped Vectashield with DRAQ5 at 5uM final concentration. Others have described the same combining DRAQ5 and ProFade Gold.
IMPORTANT: ensure that the mountant is the "native" product - DRAQ5 and DRAQ7 cannot be combined with mountants containing DAPI as the DAPI quenches the DRAQ5/DRAQ7 staining signal.
What type of instrumentation was used to collect the segmented images on this page?
An epifluorescence microscope. This type of segmentation is derived by software and can be achieved with any epifluorescent or confocal microscope, including the automated microscopy platforms used in high content screening (including GE InCell 1000,2000, 3000; Molecular Devices ImageXpress Micro, Ultra; Perkin Elmer Opera, Operetta; BD Biosciences Pathway 455, 855 and bespoke platforms).
Call 1-800-829-3194 for technical assistance or email Technical Support: help {at} immunochemistry.com.
DRAQ7™ is a trademark of Biostatus Ltd. DRAQ7 is intended for research purposes only.
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