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MTT Cell Proliferation Assay Kit (Colorimetric)

based on 3 citations in multiple journalsMTT Cell Proliferation Assay Kit (Colorimetric)34.1 4
Non-radioactive and high-throughput method to measure cell proliferation/viability/cytotoxicity
Catalog #: K299

Product Details

Cat # +Size K299-1000
Size 1000 assays
Detection Method Absorbance (590 nm)
Applications Measurement of cell proliferation in response to growth factors, cytokines, mitogens and nutrients.
Analysis of cytotoxic and cytostatic compounds such as anticancer drugs, toxic agents and other pharmaceuticals.
Assessment of physiological mediators that inhibit cell growth.
Features & Benefits • Non-radioactive
• High-throughput
• The entire assay can be performed directly in a 96-well plate and does not require washing/harvesting/or solubilization steps.
Kit Components • MTT Reagent
• MTT Solvent
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.


Quantification of the number of viable cells is an indispensable tool in cell biology research. BioVision’s MTT Cell Proliferation Assay Kit is a sensitive method for quantification of viable cells in proliferation and cytotoxicity assay. The method is based on the conversion of water soluble MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) compound to an insoluble formazan product. Viable cells with active metabolism convert MTT into formazan, however, dead cells lose this ability. Thus color formation serves as a useful and convenient marker of only the viable cells. The measured absorbance (590 nm) is proportional to the number of viable cells. This assay kit provides an easy-to-use, non-radioactive, and high-throughput method for cell proliferation, cell viability, chemotaxis, cytotoxicity, and apoptosis.

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Should the media in background control well be serum-free media? Also, after 3 hour incubation with MTT reagent, should I remove MTT reagent or directly add MTT solvent without removing MTT reagent?
Use serum-free media for the background control wells too. Whether to remove the MTT reagent or not depends on the type of cells you have. For adherent cells, you can remove the MTT reagent gently and then add the MTT solvent. For suspension cells, one may lose the cells while removing the MTT reagent. Thus, we do not recommend removing the MTT reagent. The MTT solvent can be added directly and the total volume will be around 250 µl in the wells.
What are the known intereferences with this assay?
A variety of chemical compounds are known to interfere with the MTT assay. These are mostly reducing compounds that lead to non-enzymatic reduction of the MTT to formazan. Examples include ascorbic acid, vitamin A, sulfhydryl-containing compounds including reduced glutathione, coenzyme A, and dithiothreitol. Certain plant extracts and polyphenolic compounds can also interfere with the MTT assay.
Can you use media containing phenol red with this assay?
It's preferred not to use phenol red.
Why do I see high absorbance from media only?
The media contaminated with cells/bacteria or it might contain ascorbic acid. Always use sterile technique to work with the reagetns . Try to work with sterile pipette tips and inside a biological hood.
Why is the MTT reagent blue/green in color?
The reagent is either contaminated by a reducing agent or cell/bacterial contamination or it received excessive exposure to light. The reagent should be discarded. New reagent should be processed under sterile conditions and kep in the dark at 4°C.
Can the kit work on bacteria or yeast cells?
The kit has been standardized for mammalian cells only
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact your regional BioVision distributor.
What is the shelf life of this kit?
 BioVisions’s assay kits expire 6 months from the date of shipment. Some components of the kit may expire sooner as mentioned in the data sheet
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
Chung TW et al., (2017) Sinularin induces DNA damage, G2/M phase arrest, and apoptosis in human hepatocellular carcinoma cells, BMC Complement Altern Med, 2017, 17(1):62
Li, Q. et al. (2017) Biological function and mechanism of miR-33a in prostate cancer survival and metastasis: via downregulating Engrailed-2, Clin Transl Oncol. 2017 May;19(5):562-570.
Dokun et al., ADAM12: a genetic modifier of preclinical peripheral arterial disease.  Am J Physiol Heart Circ Physiol, Sep 2015; 309: H790 - H803.
For more citations of this product click here