Living / Dead Viability_Cytotoxicity kit for mammalian cells

The Living/Dead Viability/Cytotoxicity Kit for Animal Cells provides a two-color fluorescence staining on both

live and dead cells using two probes that measure two recognized parameters of cell viability — intracellular

esterase activity and plasma membrane integrity. The kit is suitable for use with fluorescence microscopes,

fluorescence multiwell plate scanners and flow cytometers and other fluorescence detection systems. The

assay principles are general and applicable to most eukaryoticcell types, including adherent cells and

certain tissues, but not to bacteria or yeast. It is generally faster, less expensive, safer and a more sensitive

indicator of cytotoxic events than alternative methods. Live cells are distinguished by the presence of

ubiquitous intracellular esterase activity, determined by the enzymatic conversion of the virtually

nonfluorescent cell-permeant calcein AM to the intensely fluorescent calcein. The polyanionic dye calcein is

well retained within live cells, producing anintense uniform green fluorescence in live cells (Ex/Em ~495

nm/~520 nm). Propidium iodide (PI) enters cells with damaged membranes and undergoes a 30-fold

enhancement of fluorescence upon binding to nucleic acids, thereby producing a bright red fluorescence in

dead cells (Ex/Em ~528 nm/~617 nm). PI is excluded by the intact plasma membrane of live cells.

Applications;

Fluorescence Microscopy, Flow Cytometry