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Green Live/Dead Stain

CAT: 0436-6342Size: 50 µLDry Ice: NoHazardous: No
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CAT#:0436-6342Size:50 µL
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24/48H Stock Items & 2 to 6 Weeks non Stock Items.
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Description
Green Live/Dead Stain (catalog #6342) is a vital dye that exhibits intact cell membrane exclusion properties analogous to the popular red fluorescing vital dyes, Propidium Iodide (PI), 7-amino- actinomycin D (7-AAD) and DRAQ7™. Like the red fluorescing dyes, Green Live/Dead Stain is excluded from intact, healthy cells due to its polar nature. In the presence of cells exhibiting compromised membrane integrity, 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 Green Live/Dead Stain to be used in flow cytometry-based protocols to assess the percentage of late apoptotic, necrotic, and membrane-compromised cells within a sample cell population. When bound to nucleic acids, the maximum absorption of Green Live/Dead Stain is 495 nm and the maximum emission is 512 nm (Figure 1). Cells can be viewed through a fluorescence microscope (Figures 2 & 5) or analyzed with a flow cytometer (Figure 4). Green Live/Dead Stain is provided as a 500 µM concentrated stock solution dissolved in DMSO. For flow cytometry applications, a staining concentration of 50 nM is recommended. Therefore, using sample sizes of 0.5 mL, a single 50 µL vial (catalog #6342) provides enough reagent for 1000 tests. For fluorescence microscopy applications, a usage concentration of 0.5 µM is suggested. In this way, a vial is sufficient for 100 tests (0.45 mL sample sizes). Green Live/ Dead Stain can be used with ICT’s red FLICA® 660 caspase inhibitor reagents (e.g., catalog #9120) to identify four populations of cells: living; early apoptotic; late apoptotic; and necrotic (Figures 4 & 5).
Target
Dead cells, necrosis
Label
ICT
Cell Type
Jurkat
Type
Cell Viabiity
Detection Method
Flow cytometry, Fluorescence microscope
Wavelength
495 nm / >512 nm, when bound to nucleic acids
Assay Protocol
1. Allow product to equilibrate to room temperature (25°C) prior to use., 2. Add stop solution to plate Well. Equal volumes of TMB microWell substrate and stop solution should be used. We recommend using 100 µL of TMB substrate and 100 µL of Stop Solution for TMB Substrates per Well. The stopped TMB reaction product is stable for 1 hour., 3. Read absorbance at 450 nm within 60 minutes. For best results, sample absorbance should be monitored and stopped before values exceed 2.0 OD units.
Sample Type
Cell culture
Concentration
500 µM
Shipping Conditions
Ships overnight (domestic), International Priority Shipping
Storage Temperature
-20°C
Notes
20% discount
Cellular Imaging & Detection
Cellular Imaging
Target Description
Green Live/Dead Stain is a membrane impermeant fluorescent vital stain for differentiating live and dead cells., , This green Live/Dead Stain is a 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, Green Live/Dead Stain is excluded from intact, healthy cells due to its polar nature. In the presence of cells exhibiting compromised membrane integrity, 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 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. The green fluorescence emission permits new combinations of dyes to be used in the understanding of cell death and apoptosis, such as use with red SR-FLICA® assays, far-red FLICA 660 caspase assays, and other visible range fluor-tagged ligands., , Green Live/Dead Stain only stains nuclei in dead and permeabilized cells. Green Live/Dead Stain does not enter intact, live cells. Cells may be fixed or permeabilized for use of Green Live/Dead Stain as a nuclear counterstain. With negligible toxicity, Green Live/Dead Stain can be used in long-term viability assays to assess cytotoxicity and apoptosis., , For microscopy applications, 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.

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