Table of Contents
How does DRAQ5 work?
DRAQ5™ is a far-red emitting, anthraquinone compound that stains nuclei in live cells. It is permeant to live cells and thus can be used for cell cycle analysis and/or staining of nucleated cells. It is optimally excited at 568 nm, 633 nm, and 647nm and can be detected using 695LP, 715LP, and 780LP filters.
What does DRAQ5 stain?
DRAQ5™ Fluorescent Probe is a far-red DNA stain for fluorescent cellular imaging applications with live or fixed cells. Because of its far-red excitation and emission, the DRAQ5 Stain can be multiplexed with many other fluorophores.
What does DRAQ5 bind?
At higher concentrations (3 and 7.5 microM), DRAQ5 interfered with binding of H2B core histones to DNA.
Is DRAQ5 toxic?
The DRAQ5 is toxic in the sense that it binds DNA and interferes with replication: cells appear to remain viable but be halted in G2/M phase. So if you are after culturing your cells after analysis, then DRAQ5 is not for you.
Does DRAQ5 stain dead cells?
DRAQ5™ is an anthraquinone dye with high affinity for double-stranded DNA. It is a membrane-permeable dye that can label live or fixed/dead cells. In flow cytometry, this dye can be used to distinguish nucleated and non-nucleated cells.
Can Hoechst be used for live cell imaging?
Dyes that bind to DNA, such as Hoechst 33342, are commonly used to visualize chromatin in live cells by fluorescence microscopy.
What is phototoxicity in microscopy?
Phototoxicity is the process by which upon illumination, with high laser power or for prolonged periods, the imaged organisms/cells are damaged. Phototoxicity can cause cellular membrane blebbing, vacuole formation and even cell death.
What is DAPI and when is it used?
A simple-to-use fluorescent stain, 4′,6-diamidino-2-phenylindole (DAPI), visualizes nuclear DNA in both living and fixed cells. DAPI staining was used to determine the number of nuclei and to assess gross cell morphology. Following light microscopic analyses, the stained cells were processed for electron microscopy.
Can you use DAPI on live cells?
It is used extensively in fluorescence microscopy. As DAPI can pass through an intact cell membrane, it can be used to stain both live and fixed cells, though it passes through the membrane less efficiently in live cells and therefore provides a marker for membrane viability.
Is Hoechst better than DAPI?
Hoechst dyes are generally preferred for live cell staining over DAPI because they are less toxic and more cell permeant.
How do you test for phototoxicity?
Although specimens show diverse signs of light-induced damage, common themes do exist. A frequent recommendation for assessing phototoxicity is to look for tell- tale morphological signs such as cellular swelling and rounding, blebbing, or the appearance of vacuoles.
What wavelength is DAPI?
Fluorescence properties When bound to double-stranded DNA, DAPI has an absorption maximum at a wavelength of 358 nm (ultraviolet) and its emission maximum is at 461 nm (blue). Therefore, for fluorescence microscopy, DAPI is excited with ultraviolet light and is detected through a blue/cyan filter.
Why is DAPI used in staining?
DAPI is generally used to stain fixed cells since the dye is cell impermeant, although the stain will enter live cells when used at higher concentrations.
What is the protocol for surface staining with DRAQ5?
Product Details 1 Perform surface staining following protocol of choice. 2 Wash cells twice with phosphate buffered saline (PBS). Sodium azide interferes with DRAQ5™ staining, thus it is recommended to stain in PBS (without calcium, magnesium, or sodium azide) or 3 Dilute DRAQ5™ to required concentration. Weitere Artikel…
What is DRAQ5 fluorescent probe?
DRAQ5 Fluorescent Probe emits in the far-red region, is lipophilic and crosses cell and nuclear membranes in live and fixed cells and tissues for rapid DNA staining. This stain is water-soluble, supplied ready to use and does not require RNase, cell lysis, or a washing step, making it compatible with automation.
What is DRAQ5™?
DRAQ5™ is a far-red emitting, anthraquinone compound that stains nuclei in live cells. It is permeant to live cells and thus can be used for cell cycle analysis and/or staining of nucleated cells. It is optimally excited at 568 nm, 633 nm, and 647nm and can be detected using 695LP, 715LP, and 780LP filters.
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.