Bitan lab wiki: Cell viability by test by MTT assay

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Cell viability by MTT assay

Cell viability was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell-metabolism assay

Reagents and accessories:

MTT dye (Sigma), stop solution and solubilizing solution (Invitrogen), White plate (Costar)


Cells were treated with freshly prepared peptide for desire time.

Following treatment, 15 μl of MTT was added to each well and incubated for 4 h at 37ºC.

Then, stop solution was added and kept overnight at room temperature.

Next day the OD was measured using a Synergy plate reader (Bio-TEK instruments, Winooski, VT) at 540 nm.

The cell viability results were normalized to the medium control group and expressed as mean ± SEM. The cell viability was calculated by the following formula:

Cell viability (% of control) = {1-(OD of test sample-OD of full killed sample)/(OD of full killed sample-OD of medium control sample)}* 100