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Antibodies

Green Live/Dead Stain

Product Sizes
50 ul
6342-50UL
About this Product
SKU:
6342
Application:
Flow Cytometry
Concentration:
500 uM
Extra Details:
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 uM 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 uL vial (catalog #6342) provides enough reagent for 1000 tests. For fluorescence microscopy applications, a usage concentration of 0.5 uM 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).
Immunogen:
Dead cells, necrosis
Physical State:
Cell Culture
Shipping Conditions:
Blue Ice
Storage Conditions:
-20[o]C
Type:
Antibody: Stains & Enhancers
Figure 1. Fluorescence spectra
Figure 2. Jurkat cells were exposed to 3% formaldehyde for 30 minutes. Following the formaldhehyde treatment, cells were stained with Green Live/ Dead stain and then imaged with a Nikon E800 microscope (DIC overlay shown). Cells with compromised membranes stained green, while cells with intact membranes excluded the Green Live/Dead stain and remained unstained. Data courtesy of Mrs. Tracy Murphy, ICT.
Figure 3. Jurkat cells were transferred to a suboptimal culture environment (serum-free DMEM at RT and 0.03% CO2). Viability results obtained with Green Live/Dead Stain after 3 hours were consistent with the commonly used vital dyes PI, 7-AAD, and DRAQ7.
Figure 4. Staurosporine-treated Jurkat cells were stained with FLICA® 660 Poly Caspase Assay Kit (Cat. 9120) and Green Live/Dead Stain, then analyzed by flow cytometry. Four populations of cells were detected: (LL) unstained live cells; (LR) green necrotic cells; (UR) late apoptotic cells stained with red FLICA® 660 and Green Live/Dead Stain; and (UL) red early apoptotic cells. The majority of treated cells (73.2%) are in early apoptosis (bottom plot, UL). Data courtesy of Mrs. T. Murphy, ICT
Figure 5. Green Live/Dead Stain and FLICA® 660 Poly Caspase detection reagent (Cat. 9120) were used to assess cell death in staurosporine-treated Jurkat cells. Cells were imaged with a Nikon E800 microscope. The majority of cells were in the early stages of apoptosis (active caspases stained red with FLICA® 660). One cell in this field of view (bottom right corner) exhibited both red and green fluorescence, indicating the loss of membrane integrity, characteristic of late-stage apoptosis.