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DRAQ5™: Technical Details

• Compound code:                         DRAQ5™

• Molecular Weight:                         412.54

• State of original compound:       Dark blue crystalline solid

• Stable at room temperature.

• Suggested storage at 4ºC and under light protection.

• Do not freeze the solution.

DRAQ5™: Sample preparation and analysis

1. Consider safe handling procedures (see material safety data sheet).  Regard the agent as a DNA binding chemical with potentially mutagenic properties as with other DNA intercalators.

2. To use DRAQ5™ with living cells, add at concentrations of 1-20 µM (preferably at 10 or 20 µM for cells at a density of 4 x 10⁵ml) by direct addition to culture medium or buffer.  DNA staining occurs at room temperature but is very rapid (approx 1-3 min) at 37ºC.

3. Flow cytometry applications can use the standard 488 nm or 633 nm excitation modes and filter configurations available on bench-top instruments, or 647 nm excitation available on research cytometers.  Suggested optimal emission filters are 695LP, 715LP or 780 LP.  Combining DRAQ5™ with FITC does not require fluorescence compensation unlike applications using propidium iodide.  Whole blood preparations can be stained directly using DRAQ5™.  As with all DNA stains they tend to contaminate the sample tubing.  In the case of DRAQ5™ simple rinsing appears to clear the sample tube of any dye that may be released into the following sample.  With repeated use the dye may stain the sample tube light blue but this does not produce a problem and we have never observed cross contamination of samples.  A 400 µl staining reaction at 20 µM DRAQ5™ permits 125 assays for each vial supplied.

4. This DRAQ™ dye can be used as a DNA stain for living cells (or fixed cells) which upon equilibrium can be used to measure cell cycle distribution or nuclear integrity.  It is not recommended for viable cell sorting since the persistence of DRAQ5™ on cellular DNA is ultimately cytotoxic.

5. For imaging as a marker for cell nuclei (using laser excitation of 568 nm, 633 nm or 647 nm wavelengths) use at a concentration of 1-20 µM. Cells can be mounted directly in DRAQ5™ dye and imaged immediately.  A 200 µl staining reaction at 5 µM DRAQ5™ permits 1000 assays for each vial supplied.

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