When I perform the MTT assay on BMDMs, the unstimulated cells show lower absorbance values, around 0.2–0.3 at 570 nm. What could be the reason for this? The assay was performed in a 96-well plate with 1 × 10⁵ cells per well.
Malcolm Nobre
Hello Lara Parreiras,
Unstimulated BMDMs, which are not actively responding to stimuli, typically have lower metabolic activity compared to stimulated or activated BMDMs.
MTT assay relies on the reduction of MTT by mitochondrial dehydrogenases in metabolically active cells, which produces a colored formazan product. Therefore, cells with low metabolic activity will produce less formazan, leading to lower absorbance readings in MTT assay.
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