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XTT Cell Viability Assay Kit

A simple, colorimetric assay for determining live cell numbers by absorbance on a microplate reader.
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1,000 assays
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Product Description

The XTT Cell Viability Assay Kit provides a simple method for determining live cell numbers by absorbance on a microplate reader. XTT is a tetrazolium derivative that is turned into a water-soluble orange product after reduction by mitochondrial enzymes that are only present in metabolically active live cells. The amount of orange product generated is proportional to the number of living cells in the sample.

  • Measure cell metabolism by reading absorbance at 475 nm
  • Non-radioactive, simple procedure
  • XTT is a tetrazolium derivative
  • Endpoint assay
  • Unlike MTT, does not require solubilization step

Determination of live cell numbers is often used to assess the rate of cell proliferation and cytotoxicity caused by drugs and cytotoxic agents. The non-radioactive MTT assay developed by Mossman is one of the most popular viability assays; however, the product produced is non-soluble, and requires a solubilization step. Unlike MTT, XTT is reduced to a highly water-soluble orange-colored product instead of the insoluble formazan from MTT, thus eliminating the solubilization step required for the MTT assay.

The amount of water-soluble product generated from XTT reduction is proportional to the number of living cells in the sample and can be photometrically quantified at 475 nm. Continuous color development permits measurements at multiple time points for extended detection range.

Product Attributes

Size
1,000 assays
Apoptosis/viability marker
Metabolic activity
For live or fixed cells
For live/intact cells
Assay type/options
Endpoint assay, High-throughput assay, Homogeneous assay, Real-time imaging
Detection method/readout
Microplate reader (absorbance)
Storage Conditions
Store at 2 to 8 °C

Documents, Protocols, SDS and COA

References

1. PLoS ONE (2013) 8(8): e71165 doi:10.1371/journal.pone.0071165
2. ACS Nano (2013) 7, 9, 7524-7533 doi.org/10.1021/nn403722x
3. Oncogene (2014) 6 October doi:10.1038/onc.2014.328
4. J Biol Chem (2014) Sep 26;289(39):26882-94. doi: 10.1074/jbc.M114.567685
5. Environ. Sci Nano (2015) 2, 370-379, doi: 10.1039/C5EN00031A
6. Lab Invest (2015) Mar;95(3):296-307 doi: 10.1038/labinvest.2014.161
7. Mol Biol Rep (2016) 43, 229–240 doi:10.1007/s11033-016-3948-4
8. Nat Commun (2016) 7, 11551 doi:10.1038/ncomms11551

Citations

1. PLoS ONE (2013) 8(8): e71165 doi:10.1371/journal.pone.0071165
2. ACS Nano (2013) 7, 9, 7524-7533 doi.org/10.1021/nn403722x
3. Oncogene (2014) 6 October doi:10.1038/onc.2014.328
4. J Biol Chem (2014) Sep 26;289(39):26882-94. doi: 10.1074/jbc.M114.567685
5. Environ. Sci Nano (2015) 2, 370-379, doi: 10.1039/C5EN00031A
6. Lab Invest (2015) Mar;95(3):296-307 doi: 10.1038/labinvest.2014.161
7. Mol Biol Rep (2016) 43, 229–240 doi:10.1007/s11033-016-3948-4
8. Nat Commun (2016) 7, 11551 doi:10.1038/ncomms11551

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FAQs

Apoptosis, Necrosis, and Cell Viability Kits

It has been reported in publications that concentrations of serum above 10% in the assay may affect the results.

See the following publications for more information

https://doi.org/10.2144/04373ST05

https://dx.doi.org/10.1007%2Fs10616-007-9057-4

Our ViaFluor® SE Cell Proliferation assay is a dye dilution assay for cell division, like CFSE and CellTrace™ Violet from Thermo. This type of assay is commonly used to measure lymphocyte proliferative responses in culture and in vivo (if the labeled cells are injected back into mice). It requires flow cytometry to analyze and allows you to count how many cell divisions have occurred in the labeled cells.

For more information and a typical procedure for using fluorescent ViaFluor® SE Dyes with PMBCs, which can easily be adapted for use with other cell types, please see our Tech Tip: Measuring Cell Division in PMBCs by Flow Cytometry

If flow cytometry is not an option, we offer absorbance-based and fluorescence-based microplate assays for quantitating cell numbers. These measure mitochondrial activity (resazurin/MTT/XTT) or intracellular esterase activity (calcein AM) as a readout of viable cell numbers. Please visit the Cell Viability and Apoptosis technology page for more information.

The ATP-Glo™ assay is a luminescence assay for cellular ATP levels, which are proportional to the number of live cells. This assay requires a luminometer.

CellTrace is a trademark of Thermo Fisher Scientific.

Our Resazurin Cell Viability Assay (Cat. No. 30025) has red fluorescence (Ex/Em 530-560/590 nm), and is specifically designed for microplate reader. It is an economical, easy-to-use, and homogeneous (no-wash) assay for quantifying live cells. It is similar to alamarBlue®, PrestoBlue®, and CellTiter-Blue®.

The Calcein AM Cell Viability Assay (Cat. No. 30026) has green fluorescence (Ex/Em 485/530 nm), and also works well for microplate reader. This assay requires culture medium to be removed from cells before adding the viability dye in buffer. We also offer the Viability/Cytotoxicity Assay for Animal Live & Dead Cells, which combines calcein-AM with the fluorescent dead cell stain EthD-III, and is compatible with microplate reader.

alamarBlue is a registered trademark of Trek Biosystems. CellTiter-Blue is a registered trademark of Promega Corporation. PrestoBlue is a registered trademark of Thermo Fisher Scientific.
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