Bitan lab wiki: Cell viability by test by MTT assay

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)

Protocol:

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