Sample Protocol
Sample Protocol: Typical use: Flow Cytometry 1000 assays - 5 ul of working solution in 495 ul cells (end concentration 50 nM) Imaging 100 assays - 50 ul of working solution in 450 ul cells (end concentration 500 nM) Example Protocol for Flow Cytometry: Spike samples with the Green Live/Dead Stain working solution. Incubate samples approximately 10 min at RT, protected from light. Analyze with a flow cytometer using a blue laser at 488 nm and a 530/30 (FL1) emission filter setting, or similar. Example Protocol for Microscopy: Spike samples with the Green Live/Dead Stain working solution. Incubate samples approximately 10 min at RT, protected from light. When bound to DNA, the peak absorption of CellMetrix Green Live/Dead Stain is 495 nm and the maximum emission is 512 nm. Visualize with a fluorescence microscope using optical filters that best approximate these settings.
Target
Dead Cells, Necrosis
495 nm / >512 nm
When bound to nucleic acids, the maximum absorption of CellMetrix Green Live/Dead Stain is 495 nm and the maximum emission is 512 nm. Cells can be viewed through a fluorescence microscope or analyzed with a flow cytometer.
Excitation / Emission
Multi-wavelength imaging with UV / vis fluorochromes
Fluorescence enhancement upon dsDNA binding
Compatible with optics of flow, laser scanning cytometers and confocal and lamp-based fluorescence microscopes
Other Notes
Small volumes of CellMetrix Green Live/Dead dye vial(s) may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container`s cap. Certain products may require to ship with dry ice and additional dry ice fee may apply.
Related Product Information for
CellMetrix Green Live/Dead dye
Description: CellMetrix Green Live/Dead Stain is a new live-cell impermeant, green fluorescence-emitting DNA dye for viability, apoptosis and necrosis studies, and fixed cell nuclear counterstaining. Rapid staining of necrotic or permeabilized cells. Binds to dsDNA/nuclei of DEAD or permeabilized cells. Combination with live cell dyes for live/dead discrimination. Easy to use - no lyse, no wash, no RNase needed. Ideal for use with red SR-FLICA and FLICA 660 caspase assays.
Background: CellMetrix Green Live/Dead Stain is a new live/dead DNA dye for apoptosis and necrosis studies. CellMetrix Green Live/Dead Stain is a membrane impermeant DNA dye for the rapid and convenient staining of the nuclear DNA of dead or fixed cells. This new green DNA dye is an ideal live/dead stain to assess necrosis in cytotoxicity studies, as well as a reliable DNA dye in fixed cell samples with immunofluorescence, immunohistochemistry, and high content screening applications. This green Live/Dead Stain is a recently developed vital dye that exhibits intact cell membrane exclusion properties analogous to the popular red fluorescing vital dyes, Propidium Iodide (PI), 7-aminoactinomycinD (7-AAD), and DRAQ7. Like the red fluorescing DNA dyes, CellMetrix Green Live/Dead Stain is excluded from intact, healthy cells due to its polar nature. In the presence of cells exhibiting compromised membrane integrity, CellMetrix Green Live/Dead Stain penetrates the cell and nuclear membrane barriers and intercalates tightly to DNA in a manner analogous to PI and 7-AAD. When bound to DNA, it acquires a greatly enhanced fluorescence potential (>2000X) in the green emission range. These important vital dye properties enable CellMetrix Green Live/Dead Stain to be used in flow cytometry-based protocols and microscopy analysis techniques to assess the percentage of late apoptotic, necrotic, and membrane-compromised cells within a sample cell population. CellMetrix Green Live/Dead Stain is supplied as a concentrated stock solution. Each 50 ul vial is sufficient for 1000 flow cytometry tests or 100 microtiter plate wells per our sample protocol. Optimal Ex/ Em: 495 nm/ 512nm Identification of Dead Cells / Membrane-Compromised Cells: Live Cell Membrane-Impermeant CellMetrix Green Live/Dead Stain only stains nuclei in dead and permeabilized cells. CellMetrix Green Live/Dead Stain does not enter intact, live cells. Cells may be fixed or permeabilized for use of CellMetrix Green Live/Dead Stain as a nuclear counterstain. Green emission The green fluorescence emission (512 nm optimal) of CellMetrix Green Live/Dead Stain permits new combinations of dyes to be used in the understanding of cell death and apoptosis. CellMetrix Green Live/Dead Stain is ideal for use with red SR-FLICA assays, far-red FLICA 660 caspase assays, and other visible range fluor-tagged ligands. Non-toxic in long-term culture With negligible toxicity, CellMetrix Green Live/Dead Stain can be used in long-term viability assays to assess cytotoxicity and apoptosis. Nuclear Counterstaining with Fixed Cells & Tissue Sections: For microscopy applications, CellMetrix Green Live/Dead Stain will function as a DNA counterstain for fixed cell and tissue samples in IF, IHC, high content screening, and cell-based assays. Visible Range Excitation: Unlike blue fluorescence DNA dyes, CellMetrix Green Live/Dead Stain is excited with a blue laser in a flow cytometer and blue excitation filters in imaging instruments. Make compound screens more effective: Visible range excitation avoids a "white-out" effect from coincidental UV fluorescence of compounds when using DAPI or Hoechst DNA dyes. Spectrally compatible with UV excited fluors Compatible with PE-tagged probes, SR-FLICA, or FLICA 660 caspase assays.
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