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A simple, colorimetric assay for determining live cell numbers by absorbance on a microplate reader.
The MTT Cell Viability Assay Kit provides a simple method for determining live cell numbers by absorbance on a microplate reader. MTT is a tetrazolium salt that is turned into a purple formazan product after reduction by mitochondrial enzymes that are only present in metabolically active live cells. The amount of formazan product generated is proportional to the number of living cells in the sample.
Determination of live cell numbers is often used to assess the rate of cell proliferation and cytotoxicity caused by drugs and cytotoxic agents. Among all non-radioactive viability assays, MTT assay developed by Mossman is one of the most versatile and popular assays. At the end of the assay, the cells containing the formazan product are solubilized and then photometrically quantified at 570 nm.
Also see the XTT Cell Viability Assay Kit, which does not require solubilization before absorbance measurement.
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8. Nat Commun (2016) Feb 16;7:10686. doi: 10.1038/ncomms10686
1. PLoS ONE (2011) 6(8): e22635. doi:10.1371/journal.pone.0022635
2. Cell (2011) 145, 941-955. doi:10.1016/j.cell.2011.05.004
3. PLoS ONE (2012) 7(4): e35866. doi:10.1371/journal.pone.0035866
4. J. Virol (2013) Jul;87(14):8179-94. doi:10.1128/JVI.00443-13
5. J Am Chem Soc. (2014) Apr 9;136(14):5295-300. doi: 10.1021/ja408505n.
6. Endocrinology. (2015) Dec;156(12):4695-706. doi: 10.1210/en.2015-1500
7. J Mol Neurosci. (2015) May;56(1):122-30. doi: 10.1007/s12031-014-0473-6
8. Nat Commun (2016) Feb 16;7:10686. doi: 10.1038/ncomms10686
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
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.