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Viability/Cytotoxicity Assay Kit for Animal Live & Dead Cells

The Viability/Cytotoxicity Assay Kit for Animal Live & Dead Cells provides green/red fluorescent staining of live and dead cells, respectively. The assay employs two probes that detect intracellular esterase activity in live cells and compromised plasma membrane integrity in dead cells.

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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

Endpoint assay, No-wash staining, Real-time imaging, Homogenous assay

Colors

Green, Red

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150 assays
300 assays
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Product Description

The Viability/Cytotoxicity Assay Kit for Animal Live & Dead Cells provides green/red fluorescent staining of live and dead cells, respectively. 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 (#40050 and #40051) 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 thus brighter fluorescence (30% brighter than EthD-I).

Spectral Properties
Calcein (end product after hydrolysis of calcein AM):
Ex/Em: 494/517 nm (pH 8)

Ethidium homodimer III (EthDIII):
Ex/Em: 530/620 nm (with DNA)

References

1. Biotechniques 29, 874-880 (2000).

 

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Support & Faq

Apoptosis, Necrosis, and Cell Viability Kits

Fixation is not recommended after staining with the Viability/Cytotoxicity Assay Kit (cat. # 30027) as the dead cell dye, EthD-III will transfer to all cells and not maintain dead cell-specific staining.

Viability qPCR (vPCR) can be an alternative strategy. Using dead cell-specific dyes, PMA (cat. # 40019) or PMAxx (cat. # 40069) that covalently modify DNA (of only dead cells) after photo-crosslinking, a simple quantitative PCR (qPCR) amplification is used to selectively amplify live-cell DNA. Learn more about the vPCR technology.

Category: Apoptosis, Necrosis, and Cell Viability Kits

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The ViaFluor® SE dyes are susceptible to hydrolysis. Ideally the DMSO stock solution should be prepared on the day of use, with the intent that each vial supplied is for single use. If smaller aliquots are desired, it would be best to aliquot the initial DMSO stock, at the time of reconstitution, into single use quantities and store at -20C, protected from light and moisture (desiccated), for up to a month. However, activity may be reduced over time. The dyes should only be added to aqueous buffer immediately before staining. The aqueous solution cannot be stored for reuse.

Category: Apoptosis, Necrosis, and Cell Viability Kits

← FAQs

MTT and XTT are colorimetric based assays, while resazurin can be measured using colorimetric or fluorescence detection. MTT is not a soluble product, so the cells must be lysed to solubilize the formazan salt before absorbance can be measured. XTT and resazurin do not require cell lysis, allowing kinetic monitoring of the same samples at different timepoints.

Category: Apoptosis, Necrosis, and Cell Viability Kits

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Shipping/Shelf-life

Shipping & Handling Disclaimer: Shipping and handling methods are calculated based on item temperature storage recommendation. Large, multi-item orders may be assessed additional fees. Shipping to Puerto Rico may be assessed additional fees. A Biotium customer service representative will contact you if additional fees are required. For expedited shipping, please request in the Order Notes section of the checkout page. Please note that products with recommended storage at 4°C or -20°C may ship at ambient temperature. This will not affect product performance. When you receive the product, place it under the recommended storage conditions.