MTT Assay (by BCRC)

( www.abnova.com ) - Bioresource Collection and Research (BCRC) provided this AbAbVideo to introduce the protocol of MTT assay. MTT assay allows assessing the viability and the proliferation of cells. This is a colorimetric assay that measures the reduction of yellow 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) by mitochondrial succinate dehydrogenase. More videos at Abnova www.abnova.com

[@yilingl.8756]

So, I'm sorta new to this field and was wondering, what exactly does the absorbance level mean? High absorbance=...? Low absorbance = ...?

[@julesteuwen]

absorbance = -log(transmission of light)

[@kosheeka]

Absorbance is a measure of how much light is absorbed by a substance. In the context of in vitro studies, it's often used to quantify the amount of a particular molecule or compound present in a sample.
• High Absorbance generally indicates a higher concentration of the substance of interest. In MTT assay, a higher absorbance usually correlates to a greater number of viable cells.
• Low Absorbance typically indicates a lower concentration of the substance of interest. A low absorbance in MTT assay might suggest fewer viable cells or interference with the assay.

[@Ice84letters]

is it neccesary to remove the media before adding the MTT and add a media without phenol red? in my lab they disolve the formazan with isopropanol instead of DMSO, is it right?

[@kosheeka]

Yes, it's generally recommended to remove the media before adding MTT. The media can interfere with the MTT assay by absorbing or scattering light, affecting the absorbance readings. Additionally, some media components might react with the MTT reagent or the formazan product.

[@DubbedDemon]

nice

[@rayhollander]

why do you incubate for 4 hours?

[@kosheeka]

The 4-hour incubation period in the MTT assay is a balance between sufficient time for formazan product formation and minimizing cell toxicity.

[@lokkaf3526]

Poor hela cells in MTT.