Protocol Guide: MTT Assay for Cell Viability and Proliferation.
The MTT assay is a colorimetric reaction that can easily be measured from cell monolayers that have been plated in 35 mm dishes or multiwell plates.
The MTT (3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium bromide) assay, originally described by Mosmann, has been used to develop a quantitative colorimetric assay for mammalian cell survival and proliferation and the method was successively modified for better resolution (1-3).
After incubation, measure the absorbance on an ELISA plate reader with a test wavelength at 450 nm and a reference wavelength at 630 nm, and subtract the 630 nm background absorbance from the 450 nm measurement. 2 Figure 1. A linear relationship can be observed between OD.
The wavelength to measure absorbance of the formazan product is between 550 and 600 nm according to the filters available for the ELISA reader, used. The reference wavelength should be more than 650 nm. Figure 3: Proliferation of 7TD1 cells (mouse-mouse hybridoma) in response to recombinant human interleukin-6 (hIL-6) using MTT assay.
Read absorbance at 590 nm with a reference filter of 620 nm. Storage: The MTT Reagent must be kept at 4C in the dark. The Detergent Reagent can be stored at either 4C or ambient temperature. If the detergent reagent is kept at 4C, warm the bottle for 5 minutes at 37C and gently mix by inverting before use (avoid creating bubbles).
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MTT assay and SRB assay respectively. In summary, Boswellic acid and Montelukast sodium are likely to be valuable for the treatment of prostate cancer, but further studies are required for their more extensive biological evaluations. Key Words: Anti-cancer activity, Boswellic Acid, Montelukast Sodium, MTT assay, PC-3 cell line, SRB assay.