DRAQ5 Fluorescent Probe

Item Number

D266292

• Storage Temp: Store at 2-8°C,Protected from light
• Shipped In: Wet ice

Product Description

Product introduction

1. Specifications: 5mM 50 μL (suitable for 4 96-well experiments), 200 μL (suitable for 16 96-well experiments)

2. Parameters: MW= 412.49; Wavelength: Ex/Em= 646/697nm; Solvent: Water

3. Storage: 4 ℃ protected from light for 2 years

Product description

DRAQ5 is a far-infrared fluorescent living cell DNA dye with high affinity for double-stranded DNA. It is a dye that can penetrate the cell membrane and can mark live cells or fixed/dead cells. In flow cytometry, this dye can be used to distinguish between nucleated and non-nucleated cells. Because DRAQ5 can bind to DNA in a stoichiometric ratio, it can also be used to report nuclear DNA content, and is suitable for chromosome multiple and cell cycle analysis. In fluorescence microscopy analysis, it can be used as a nuclear counterstain. DRAQ5 has many applications and is highly compatible with programs widely used in existing instrument platforms, the main application areas are HCS, cell models, GFP, flow cytometry and fluorescence microscopes.

The excitation wavelength range of DRAQ5 is 488～647nm. For imaging microscopy, it is recommended to use a 633 or 647nm light source for excitation. For flow cytometry, when the dye is excited at 488nm, the 685LP dichroic mirror and 710/50 channel can be used for detection; when excited at 633nm, the 660/20 channel can be used for detection. For cell cycle/DNA analysis applications, it is recommended to use a longer wavelength filter, such as 735LP dichroic mirror and 780/60 channel to optimize the CV value of G1 and G2/M peaks. Please make sure that your instrument can detect the dye.

Due to the wide emission and excitation wavelength range of DRAQ, it is not recommended to combine DRAQ5 with other far-red fluorescent dyes that can be excited at 488 or 633 nm.

Instructions

(1) Description

In the experiment, DRAQ5 is used as the last dye to stain, because DRAQ5 does not require the remaining washing steps after staining, so DRAQ5 can be directly added to the cell-containing medium for live cell staining

(2) Operation

①. Prepare PBS buffer without sodium azide or a specific medium for specific cells.

②. Resuspend the cells in PBS or medium, and control the cell density to ≤ 4 × 105 cells/mL. For adherent cells and some tissues, roughly estimate the number of cells.

③. Add the appropriate volume of DRAQ5 staining solution of appropriate concentration according to Table 1. DRAQ5 staining solution can be directly added to the surface of tissues or adherent cells, or directly added to fresh medium

④. Mix gently, and incubate at room temperature for 5-30 minutes. Incubate at 37°C and shorten the time to 1-3min. For experiments with a long time span, such as the EGFP experiment, DRAQ5 staining solution should be added to the medium during the experiment before the agonist and antagonist are added (usually 0.5 to 3 hours), and the concentration is controlled at 1 μM. Note: If the cells have been stained with other fluorescent dyes before DRAQ5 staining, please keep away from light during the above operation.

⑤. The stained cells can be directly analyzed accordingly, without other operations such as washing. The following table shows the number of cells and the required volume and final concentration of DRAQ5.

No. of cells:	in volume:	5 µM	10 µM	20 µM
1 × 106	2500 µl	2.5 µl	5 µl	10 µl
4 × 105	1000 µl	1 µl	2 µl	4 µl
2 × 105	500 µl	0.5 µl	1 µl	2 µl
1 × 105	250 µl	0.25 µl	0.5 µl	1 µl
5 × 104	125 µl	0.13 µl	0.25 µl	0.5 µl