What is DRAQ7?
DRAQ7™ is a live-cell impermeant, far-red emitting DNA dye for viability, sample quality, apoptosis and fixed cell nuclear counterstaining. This product is useful for rapid staining of dsDNA/nuclei of dead or permeabilized cells and can be used in combination with live cell dyes for live/dead discrimination.
What does DRAQ7 stain?
DRAQ7 Dye is a bright, easy-to-use fluorescent DNA dye used to exclude non-viable cells by flow cytometry. DRAQ7 Dye is a membrane-impermeable dye that rapidly stains the double-stranded DNA (dsDNA) of dead or permeabilized cells.
Is DRAQ7 fixable?
Unlike other commonly used nuclear viability stains such as propidium iodide or DRAQ7™, NucFix™ labeling is extremely stable, allowing the cells to be fixed and permeabilized without loss of fluorescence or dye transfer between cells.
How does annexin V work?
Annexin A5 (or annexin V) is a cellular protein in the annexin group. In flow cytometry, annexin V is commonly used to detect apoptotic cells by its ability to bind to phosphatidylserine, a marker of apoptosis when it is on the outer leaflet of the plasma membrane.
What is Pi in flow cytometry?
Propidium iodide (PI) is a membrane impermeant dye that is generally excluded from viable cells. It binds to double stranded DNA by intercalating between base pairs. PI is excited at 488 nm and, with a relatively large Stokes shift, emits at a maximum wavelength of 617 nm.
What is an annexin V assay?
As a new method for detecting apoptosis, the Annexin V binding assay is based on the measurement of the loss of plasma membrane asymmetry. By simultaneously excluding nuclear dye propidium iodide (PI), apoptotic cells and necrotic cells are distinguished.
How much pi do I need for flow cytometry?
Treat the cells with ribonuclease. Add 50 µl of a 100 µg/ml stock of RNase. This will ensure only DNA, not RNA, is stained. Add 200 µl PI (from 50 µg/ml stock solution).
What does JC 1 reveal about mitochondria?
The membrane-permeant JC-1 dye is widely used in apoptosis studies to monitor mitochondrial health. JC-1 dye can be used as an indicator of mitochondrial membrane potential in a variety of cell types, including myocytes and neurons, as well as in intact tissues and isolated mitochondria.
Can I use PE and PI together?
Propidium iodide (PI) is a membrane impermeant dye that is generally excluded from viable cells. Because of these spectral characteristics, PI can be used in combination with other fluorochromes excited at 488 nm such as fluorescein isothiocyanate (FITC) and phycoerythrin (PE).
Can I fix cells after PI staining?
Propidium Iodide is a charged substance; i.e. the positively charged propidium ion does not enter living cells. If the aim is to stain all cells, then they must be fixed in ethanol before staining.
What is DRAQ7 stain used for?
DRAQ7™ is a far-red fluorescent dye that only stains the nuclei in dead and permeabilized cells and can be used in combination with common labels such as GFP or FITC. DRAQ7 is the ideal tool to study dead or membrane-compromised cells because it does not enter intact, live cells.
How do you prepare DRAQ7 for flow cytometry?
Product Usage Information. Flow Cytometry: Prepare cells for DRAQ7™ staining by resuspending cells in PBS at a concentration of no more than 5 x 105 cells/ml. For each 0.5 ml of cell suspension, dilute DRAQ7™ 1:100 by adding 5 μl to suspended cells, achieving a final concetration of 3 μM. Gently mix and incubate for 10 minutes on ice.
What is the excitation wavelength for DRAQ7?
DRAQ7™ may be excited by a wide range of wavelengths from 488-647 nm. Despite low absorbance at 488 nm, this excitation source allows for convenient combination with FITC and PE conjugates as well as EGFP. DRAQ7™ is a far-red fluorescent DNA dye that only stains the nuclei in dead and permeabilized cells.
How do you prepare for DRAQ7 emission spectroscopy?
DRAQ7 ® emission spectra when excited at various wavelengths. Flow Cytometry: Prepare cells for DRAQ7™ staining by resuspending cells in PBS at a concentration of no more than 5 x 10 5 cells/ml. For each 0.5 ml of cell suspension, dilute DRAQ7™ 1:100 by adding 5 μl to suspended cells, achieving a final concetration of 3 μM.
