Does DRAQ5 stain dead cells?

DRAQ5™ staining is accelerated at 37ºC and maybe reduced to 1-3 min. DRAQ5™ stains intact, live, fixed, permeabilized and dead cells. Cells can be analysed directly without further treatment or washing.

What is DRAQ5?

DRAQ5™ Fluorescent Probe is a far-red DNA stain for fluorescent cellular imaging applications with live or fixed cells. It is commonly chosen to counterstain nuclei and measure DNA content in cell proliferation studies involving cells expressing green fluorescent protein (GFP) fusion proteins.

What is Hoechst staining used for?

Hoechst 33342 is used for specifically staining the nuclei of living or fixed cells and tissues. This stain is commonly used in combination with 5-bromo-2′-deoxyuridine (BrdU) labeling to distinguish the compact chromatin of apoptotic nuclei, to identify replicating cells and to sort cells based on their DNA content.

How do I use DRAQ5?

Add DRAQ5™ directly as supplied to a final concentration of 5 – 20µM to the cells. Alternatively, DRAQ5™ can be added to the fresh buffer/ media prior to resuspending the cells in it. 4. Gently mix the cells and then incubate for 5 – 30 minutes at Room temperature.

Does Hoechst stain dead or live cells?

Dead cells tend to stain more brightly than live cells. In S. cerevisiae, DAPI and Hoechst preferentially stain dead cells with nuclear and cytoplasmic localization.

What is the difference between DAPI and Hoechst?

Hoechst dyes are typically used for staining DNA content in live cells due to its high cell membrane permeability. DAPI is typically used for staining DNA content in fixed cells due to its low membrane permeability.

What does DRAQ5 bind?

DRAQ5 was found to intercalate or bind to the minor groove of AT-rich regions [7,10]. In theoretical simulations, a bi-mechanistic interaction was also suggested [11]. We recently categorized DRAQ5 as a concentration dependent intercalator and a minor-groove binder [5].

What is DRAQ5 stain used for?

In contrast, the anthraquinone-based intercalator DRAQ5 is a far-red DNA stain but is toxic at the concentrations used for live-cell microscopy 3.

What is the color of Hoechst stain?

On the right, staining of nucleus of a living cell with UV light excitable Hoechst 33342 (blue) or far red light excitable SiR-Hoechst (red). In addition, mitochondria stained with MitoTracker Orange (green), Scale bar 10 µm.

How does Sir–Hoechst compare to other far-red DNA stains?

Thus, SiR–Hoechst outperforms the commercially available far-red DNA stains SYTO 61, Vybrant DyeCycle Ruby and DRAQ5 both in terms of labelling specificity as well as minimal toxicity. To further assess the applicability of SiR–Hoechst, we studied mitosis by confocal laser scanning three-dimensional (3D) time-lapse microscopy.

Can DRAQ5 be used as a viability dye?

Due to its cell permeability, this dye is useful for assessing DNA content and cell cycle but is not suitable to be used as a viability dye. In microscopy applications, DRAQ5™ is also useful as a nuclear counterstain for both live and fixed specimens.