MTT assay

Dr Engelbert Buxbaum engelbert_buxbaum at hotmail.com
Mon Feb 20 05:51:46 EST 2006


Devendra Dusane wrote:

>   I am presently working on MTT assay standardization for yeast. The
> assay says development of blue color by the dehydrogenase liberated by
> the yeast cells that cleaves MTT, in the experiment that I am conducting
> I am not able to get the blue color. 

Reference for MTT assay is Slater et al., Biochim. Biophys. Acta 77
(1963) 383. IIRC there is also a description in the methods section of
the Sigma-catalog.

MTT turns purple, not blue. Have you verified that MTT passes the yeast
cell wall? I have used it only for mammalian cells, there the method is
simple: Cells are grown in 5 mg/ml MTT solution in medium for 3-4 h, the
medium is discarded, cells briefly washed with PBS and the pigment
dissolved in 50 mM HCl in i-propanol. Photometric measurement is done at
570 nm. 


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