New Promega Assay Improves Cell Culture Cytotoxicity Results
2 Oct 2012Promega Corporation has announced the introduction of a novel cytotoxicity assay that can be used on cell cultures for up to 72 hours, with flexibility for kinetic analysis to determine the onset of cytotoxicity. The CellTox™ Green Cytotoxicity Assay uses an asymmetric cyanine dye as a fluorescent marker that is impermeable to live cells but strongly binds DNA from dead cells, providing a signal that is directly proportional to cytotoxicity.
The CellTox™ Green dye is photostable and non-toxic to mammalian cells, allowing multiple reads over an extended exposure period without compromising cytotoxicity data. Other similar dyes have limited applications due to photo-instability and cytotoxicity to mammalian cells. Also, enzymatic activity biomarker assays that measure cytotoxicity, such as LDH release, have a short biomarker half-life and can result in the underestimation of cytotoxicity if used in extended exposure protocols.
The CellTox Green dye is easily multiplexed with luminescent or other fluorescent cell-based assays with an add-mix-measure protocol to provide more informative data per well. The assay reagent is also easily automated and scalable for use in 96-well to 1536-well formats. This assay is ideal for determining the cytotoxic effects of various treatments on cells in culture following long-term exposure.
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CellTox™ Green Cytotoxicity Assay
Promega Corp.The CellTox™ Green Cytotoxicity Assay measures changes in membrane integrity that occur as a result of cell death. The assay is intended to assess cytotoxicity in cell culture after experimental manipulation. The assay system uses a proprietry asymmetric cyanine dye that is excluded from viable cells but preferentially stains the DNA from dead cells. When the dye binds DNA released from cells, its fluorescence properties are substantially enhanced. Viable cells produce no appreciable increases in fluorescence. Therefore, the fluorescence signal produced by the binding interaction with dead cell DNA is proportional to cytotoxicity. The CellTox™ Green Dye is non-toxic to cells, and the signal remains constant after exposure of 72 hours, making it ideal for determining toxic effects of treatments throughout an extended exposure or as an endpoint determination. Features - Benefits • Accurate Cytotoxicity Determination: The CellTox™ Green Dye stably binds DNA of cells that have lost membrane integrity throughout 72-hour exposure and won’t underestimate cytotoxicity. • Kinetic Cytotoxicity Measures: Measure cytotoxicity at convenient time points from the same sample well to detect onset of toxicity with no duplication of plates. • Simple and Flexible Protocols: Add assay reagent directly to cells prior to plating or with dosing media to perform kinetic cytotoxicity measurements, eliminating a reagent dispensing step, or add diluted dye directly to cell culture wells as an endpoint add-mix-measure assay. • Multiplexing-Compatible: Get more informative data per well and reduce cell culture expenses by multiplexing with fluorescent and luminescent cell-based assays in the same well with no sample manipulation. • Easily Automated: Easily scale from 96- to 1536-well plate formats with “no-addition” or “single-addition” protocols. Applications Determine cytotoxic effects of treatments on cells in culture after long-term exposure.