ToxCount™
ToxCount™ provides a two wavelength cell viability assay based on determination of live and dead cells. The assay uses fluorescent dyes that measure permeability of the plasma membrane of dying cells and the ability of living cells to retain intracellular esterases, respectively. Live cells are distinguished by the presence of ubiquitous intracellular esterase activity, determined by the enzymatic conversion of the non fluores…
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ToxCount™ provides a two wavelength cell viability assay based on determination of live and dead
cells. The assay uses fluorescent dyes that measure permeability of the plasma membrane of dying
cells and the ability of living cells to retain intracellular esterases, respectively. Live cells are distinguished by the presence of ubiquitous intracellular esterase activity, determined by the enzymatic conversion of the non fluorescent cell permeable calcein AM to the intensely fluorescent calcein. In contrast, ethidium homodimer (EthD-1) enters cells with damaged membranes and binds to nucleic acids where it emits a bright red fluorescent signal in dead cells.
ToxCount can be used on multiple fluorescence detection platforms, including multi-well plate
scanning, fluorescence microscopy or flow cytometry. The assay is also suitable for use with
many adherent cell lines, primary cell types and small organisms. This method is less expensive, safer and faster than traditional cell viability assays such as 51Cr-release, LDH release and
trypan blue exclusion. The calcein AM and EthD-1 fluorescent dyes have been used to study
apoptosis and the cytotoxic effects of numerous agents including viral proteins, oxalate, amyloid proteins and cells such as lymphocytes. The assay is ideally suited for rapid screening of
libraries of drugs and small molecules for cytotoxic effects and is particularly applicable to highcontent screening using the BlueShift IsoCyte laser scanner.
The dyes provided in ToxCount are easy to use, requiring only 30 minutes loading time. In addition,
these dyes can be used in homogeneous ‘no-wash’ protocols as they are non-fluorescent in solution.
Active Motif’s ToxCount Kit has been optimized specifically for scanning on the IsoCyte laser scanner. A detailed protocol is available for customers who wish to use the reagents for 96-well, high throughput
cytotoxicity assays with the IsoCyte and automated analysis software will rapidly count the number of live and dead cells, providing the user with an easily interpreted readout for each well.
