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Viability/Cytotoxicity Assay Kit for Animal Live & Dead Cells

A convenient assay for staining live (green) and dead (red) cells within the same cell population by flow cytometry or fluorescence microscopy.

Product Attributes

Apoptosis/viability marker

Live/dead assay

For live or fixed cells

For live/intact cells

Detection method/readout

Fluorescence microscopy, Flow cytometry

Assay type/options

Short term staining (≤24h), Endpoint assay


Green, Red

Storage Conditions

Store at 2 to 8 °C, Protect from light

Catalog #
150 assays
300 assays
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Product Description

The Viability/Cytotoxicity Assay Kit for Animal Live & Dead Cells contains two dyes (calcein AM and EthD-III) for distinguishing live and dead cells.


  • Fluorescence detection of live & dead cells
  • Calcein AM stains live cells with active esterase activity green
  • EthD-III stains dead cells with compromised membranes red
  • For fluorescence microscopy, flow cytometry, or fluorescence microplate reader

Kit Components

  • Calcein AM, 4 mM in DMSO
  • EthD-III, 2 mM in DMSO/H2O

Spectral Properties (Ex/Em)

  • Calcein AM (after hydrolysis, pH 8): 494/517 nm
  • EthD-III (with DNA): 279, 522/593 nm

The assay employs two probes that detect intracellular esterase activity in live cells and compromised plasma membrane integrity in dead cells. The esterase substrate calcein AM stains live cells green, while the membrane-impermeable DNA dye ethidium homodimer III (EthD-III) stains dead cells red. The kit is suitable for detection using fluorescence microscopy, fluorescence microplate reader, or flow cytometry. The assay principles are general and applicable to most eukaryotic cell types, including adherent cells and certain tissues, but not to bacteria or yeast. This fluorescence-based method of assessing cell viability can be used in place of trypan blue exclusion, 51Cr release, and similar methods for determining cell viability and cytotoxicity.

EthD-III is a proprietary DNA stain developed by Biotium to be a superior alternative to the spectrally similar dye ethidium homodimer I (EthD-I). EthD-III has higher DNA binding affinity and brighter fluorescence (45% brighter than EthD-I).



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