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Live or Dead™ Cell Viability Assay Kit
Green/Red Dual Fluorescence
This Live or Dead™cell viability uses two fluorogenic indicators: calcein AM for viable cells and a cell-impermeable DNA-binding dye for the cells with compromised membranes. Calcein AM is a hydrophobic compound that easily permeates intact live cells, and becomes strongly fluorescent upon hydrolysis by esterases. The hydrolysis of the non-fluorescent calcein AM by intracellular esterases generates the strongly fluorescent hydrophilic calcein that is well-retained in the cell cytoplasm. The esterase activity is proportional to the number of vial cells. The DNA-binding dye is quite polar and impermeable for viable cells that have intact membranes. It becomes fluorescent only upon binding to DNA of dead cells. Cells grown in black-walled plates can be stained and quantified in less than two hours. The assay is more robust and accurate than the other viability assays. It can be readily adapted for high-throughput assays in a wide variety of fluorescence platforms such as microplate assays, immunocytochemistry and flow cytometry. The kit provides all the essential components with an optimized assay protocol. It is suitable for proliferating and non-proliferating cells, and can be used for both suspension and adherent cells. Using 100 ul of reagents per well in a 96-well format, this kit provides sufficient reagents to perform 100 assays. Using 25 ul of reagents per well in a 384-well format, this kit provides sufficient reagents to perform 400 assays.
The Effect of Jurkat cells on Saponin induced cell death measured with Cell Meter&trade; Cell Viability Assay Kit. Jurkat cells at&nbsp;2 x10<sup>6</sup> cells/mL were treated with or without 0.5% Saponin for 5 minutes. Cells were centrifuged and the supernatant were replaced with fresh medium. 100 uL of untreated cells (A), 50 uL each of untreated and treated cells (B), 25 uL of untreated and 75 uL treated cells (C), and 100 uL of 0.5% saponin treated cells (D) were plated in a 96-well black wall/clear bottom Poly-D-lysine plate. The cells were incubated with 100 &micro;L/well of CytoCalcein&trade; Green/ Propidium Iodide dye-working solution for 1 hr at 37 &deg;C. The fluorescence intensity was measured at Ex/Em = 490/525 nm and 540/650 nm with bottom read mode using NOVOstar instrument (BMG Labtech). The ratio of 490/525 nm to 540/650 nm fluorescence intensity on live and dead cells were showed as indicated (n=6).<br>&nbsp;<br>&nbsp;
The Effect of Jurkat cells on Saponin induced cell death measured with Cell Meter&trade; Cell Viability Assay Kit. Jurkat cells at&nbsp;2 x10<sup>6</sup> cells/mL were treated with or without 0.5% Saponin for 5 minutes. Cells were centrifuged and the supernatant were replaced with fresh medium. 100 uL of untreated cells (A), 50 uL each of untreated and treated cells (B), 25 uL of untreated and 75 uL treated cells (C), and 100 uL of 0.5% saponin treated cells (D) were plated in a 96-well black wall/clear bottom Poly-D-lysine plate. The cells were incubated with 100 &micro;L/well of CytoCalcein&trade; Green/ Propidium Iodide dye-working solution for 1 hr at 37 &deg;C. The fluorescence intensity was measured at Ex/Em = 490/525 nm and 540/650 nm with bottom read mode using NOVOstar instrument (BMG Labtech). The ratio of 490/525 nm to 540/650 nm fluorescence intensity on live and dead cells were showed as indicated (n=6).<br>&nbsp;<br>&nbsp;
protocol
Protocol
CatalogSize
Price
Quantity
227601000 Tests
Price
22789200 Tests
Price
 
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12352200
Instrument settings
Flow cytometer
Excitation488 nm laser
Emission530/30 nm, 610/20 nm filter
Instrument specification(s)FITC, PE-Texas Red channel
Fluorescence microscope
ExcitationFITC filter (live), TRITC filter (dead)
EmissionFITC filter (live), TRITC filter (dead)
Recommended plateBlack wall/clear bottom
Fluorescence microplate reader
Excitation490 nm (live), 540 nm (dead)
Emission525 nm (live), 620 nm (dead)
Cutoff515 nm, 590 nm
Recommended plateSolid black
Documents
Protocol
Safety Data Sheet (Catalog 22789)
Safety Data Sheet (Catalog 22760)
Certificate of Analysis
Brochure: Cell Apoptosis & Proliferation
Brochure: Cell Viability & Proliferation
Flyer: Live or Dead™ Cell Staining Kits
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Page updated on August 5, 2024