DRAQ5™ far-red fluorescent DNA dye is a novel cellular imaging reagent for use in live cells or fixed cells in combination with other common fluors. DRAQ5™ emits in the far-red (665 nm and beyond), making it ideal for use with GFP and all visible range-fluor tagged ligands. Due to this spectral separation, the GFP and nuclear signals can be captured simultaneously. For this reason alone, screening time is halved.
DRAQ5 far-red fluorescent dye directly and stoichiometrically reports DNA content in cells, live or fixed. Stained cells can be gated via flow cytometry and sorted into isolated populations with different DNA content relating to the cell cycle, such as elevated G2/M or aneuploidy. The DNA from the cell fractions can then be used directly for population-restricted PCR amplification. Because DRAQ5 is a far-red emitting DNA dye this can be easily multiplexed with other parameters, like cell surface phenotype.
DRAQ5™ differentially stains both the nucleus and cytoplasm making it ideal for any HCS translocation (or Redistribution™ / Transfluor™) assay. The intensity thresholding for the two compartments improves an in silico “dilation” while providing morphometrics which can be used to indicate toxicity. DRAQ5™ has also been successfully used in HCS assays (e.g. GPCRs) measuring cell cycle, ploidy, or protein expression to gain speed, improve performance and efficiency.
To label intact live cells, cells can be stained directly without any fixation or permeabilization. After staining for a few minutes, no washing is necessary and cells are ready for imaging. DRAQ5 can be added to the assay medium in a live cell assay.
For labeling fixed cells and nuclei, fix and wash your cells before adding DRAQ5.
The 50 µL vial will provide 150 flow cytometry assays or 250 imaging assays. The 200 µL vial will provide 600 flow cytometry assays, 1000 confocal imaging assays, or 2000 assays in a HTS/HCS screen.
Visualization of PC12 cells expressing GFP-tagged target protein and stained with DRAQ5. Left to right: DRAQ5, GFP, Overlay.
Conditions: 3000-4000 cells in one 384 well. Cells were fixed with PFA and stained with 20µL of 2.5µM DRAQ5™ in PBS for 30 min at RT.
Image acquisition settings: standard Evotec Opera™ QEHS settings for 2 colours, 20x objective. Parallel images from the target and DRAQ5™ channels with excitations at 488 and 635nm: primary dichro 488/635, detection green: 535/60 (60), detection red: 690/50. The green GFP fluorescence was quantified with respect to each cell (DRAQ5 stain).
Biostatus is grateful to Drs. Andreas Scheel and Stefan Jaeger, Evotec AG, Hamburg for providing these images.
• Supplied at: 5 mM
• Excitation/Emission: 647 nm / 665-800 nm
• Method of analysis: Fluorescence microscope, flow cytometer
• Types of samples: Cell culture
• Storage: 2-8°C
• Shipping: Ships overnight (domestic), International Priority Shipping
Kidana K, Tatebe T, Ito K, Hara N, Kakita A, Saito T, Takatori S, Ouchi Y, Ikeuchi T, Makino M, Saido TC, Akishita M, Iwatsubo T, Hori Y, Tomita T. Loss of kallikrein-related peptidase 7 exacerbates amyloid pathology in Alzheimer’s disease model mice. EMBO Mol Med. 2018. Jan 8. pii: e8184. doi: 10.15252/emmm.201708184. [Epub ahead of print]. Full text
“… ion was 20 μM), lipopolysaccharide from _Escherichia coli_ (Imgenex, stock solution was dissolved in distilled water, final concentration was 1 μg/ml), and DRAQ5™ far‐red fluorescent DNA dye (ImmunoChemistry Technologies, final concentration was 1:1,000 dilution). Memantine (stock solution was dissolved in distilled water, final concentration was 30 μM) was provided by Daiichi‐Sankyo C…”