DRAQ5™ is a cell-permeable, far-red fluorescent DNA dye that can be used for labeling DNA in both live/non-fixed and fixed cells, compatible with common fluorescent labels like GFP or FITC.
Application | Reactivity | Dilution info | Notes |
---|---|---|---|
Application ICC/IF | Reactivity Reacts | Dilution info 1/1000 | Notes For Cell-based assays, Immunofluorescence microscopy and In-Cell WB = 5µM. It is highly recommended that the concentration and labelling conditions are carefully determined by each investigator for optimal performance in the assay of interest. For more specific information about the applications, please refer to the Protocol Booklet section. |
Application Flow Cyt | Reactivity Reacts | Dilution info 1/250.00000 - 1/1000.00000 | Notes For Gating of Nucleated cells = 5µM; For Cell Cycle Analysis = 20µM |
Application Fluorescence Microscopy | Reactivity Reacts | Dilution info - | Notes - |
DRAQ5™ is a cell-permeable, far-red fluorescent DNA dye that can be used for labeling DNA in both live/non-fixed and fixed cells, compatible with common fluorescent labels like GFP or FITC.
Constituents: 100% 1,5-BIS{[2- (DIMETHYLAMINO) ETHYL]AMINO}-4,8- DIHYDROXYANTHRACE NE-9,10-DIONE
DRAQ5™ is a cell permeable far-red fluorescent DNA dye that can be used in fixed or non-fixed/ live cells in combination with common labels such as GFP or FITC.
As with any cell-permeant DNA intercalating probe, DRAQ5 may inhibit cell division in long-term assays and should be tested for any effect.
DRAQ5 staining can be used in flow cytometry, live cell imaging and cell-based assays and the dye is highly compatible with standard protocols across many instrumentation platforms.
The chemical name of DRAQ5 is 1, 5-bis{[2-(di-methylamino)ethyl]amino}-4, 8-dihydroxyanthracene-9, 10-dione.
The advantages of DRAQ5 staining include
- convenient ready-to-use aqueous solution
- rapid uptake into living cells, providing a high level of nuclear discrimination
- no photobleaching effect
- can be used in most cell types, eukaryotic and prokaryotic: mammalian, bacterial, parasitic, plant, etc.
- no compensation needed with common FITC/GFP + PE combinations in flow cytometry
- no RNase treatment required
- no fluorescence enhancement upon DNA binding
- compatible with optics of benchtop flow, laser scanning cytometers and non-UV laser scanning and lamp-based confocal microscopes
SPECTRAL PROPERTIES:
Excitation
Emission (instrument dependent):
- 665 nm to infra-red max 681 nm / 697 nm intercalated with dsDNA)
- minimal overlap with vis range e.g. GFP and FITC
- Em. filters may include 695L, 715LP or 780 LP
Concentration: 5 mM
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Full details and terms and conditions can be found here:
Terms & Conditions.
HeLa cells were stained with Lamin B1 antibody - Nuclear Envelope Marker (Anti-Lamin B1 antibody - Nuclear Envelope Marker ab16048) and alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291). The cells were 100% methanol fixed (5 min) and then incubated in 1% BSA in 0.1% PBS-Tween for 1h to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the primary antibodies (Anti-Lamin B1 antibody - Nuclear Envelope Marker ab16048 & Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1μg/ml overnight at 4C. The secondary antibodies were Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (DyLight® 488), pre-adsorbed (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) (green) and Goat polyclonal Secondary Antibody to Mouse IgG - H&L (DyLight® 594), pre-adsorbed (Goat Anti-Mouse IgG H&L (DyLight® 594) preadsorbed ab96881) (red) used at 1/250 dilution for 1h at room temperature. 5μM DRAQ5 was added to the secondary antibody mixture to label nuclear DNA (pseudocolor purple).
HeLa cells were stained with beta Catenin antibody (Anti-beta Catenin antibody ab16051) and alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291). The cells were 100% methanol fixed (5 min) and then incubated in 1% BSA in 0.1% PBS-Tween for 1h to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the primary antibodies (Anti-beta Catenin antibody ab16051 & ab7921) at 1μg/ml overnight at 4C. The secondary antibodies were Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (DyLight® 488), pre-adsorbed (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) (green) and Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (DyLight® 594), pre-adsorbed (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) (red) used at 1/250 dilution for 1h at room temperature. 5μM DRAQ5 was added to the secondary antibody mixture to label nuclear DNA (pseudocolor orange).
DRAQ5™-stained nuclei in a adult Drosophila brain.
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