I am performing the MTT and I was wondering if it is necessary to have a blank if you are going to remove the medium before adding a formazan solubilising solution? Will it affect the accuracy of your results?
Hi Luveni Sonka,
Usually, you would have obtained a line of best fit using MTT on a known number of cells (500, 1000, 2000, etc...). In order to make your results comparable to this best fit to determine cell number, you will need to subtract blank values from your total value as readings may vary between readings.
Hi Luveni,
Yes , it is necessary to perform the assay with untreated cells, as a control. The background absorbance values from the control wells corresponding to nontreated cells must be subtracted from the treated cells values, in order to obtain an accurate result of the effect of a treatment on cell proliferation.
Hello Luveni
I agree with Maria. If you perform the MTT assay without the blank the OD readings obtained for the treated cells will not be accurate, and you will not be able to know the exact effect of the compound on the cells. Therefore the inclusion of blank in the assay becomes very much essential to exclude background
signal.
Regards.
Hii Luveni !!
In every experiment we use positive control and negative control so that we can compare the obtained result. In MTT assay it is necessary to add a blank, later it will help to get appropriate effect of the drug by substracting OD of treated with control.
You should have a blank. It can affect your result. One with MTT and one with simply cells
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