Hello everyone!
I measured the protein concentration by BCA for each well of HeLa cells seeded in 6-well plate (750000 cells / 2ml DMEM per well). Protein concentration for each well was approximately 400-500 µg. After same cells were exposed to DEHP, cytotoxicity was measured at the end of 24 and 48 hours of exposure as well.
Now i'm wondering that how should i correlate my results from MTT test with the results of the protein quantification?
Thank you in advance!
The results of the MTT assay are typically expressed as the optical density (OD) of the formazan product, which is directly proportional to the number of viable cells in the culture. To correlate the results of the MTT test with the protein quantification, you can consider the following points:
- The MTT assay measures metabolic activity, while the protein quantification assesses the total protein content. An increase in metabolic activity may not always directly correlate with an increase in protein content, as it depends on various factors such as cell size, organelle content, and metabolic state.
- When correlating the MTT assay results with protein quantification, it's important to consider that the MTT assay provides a relative measure of cell viability and proliferation, while protein quantification gives the absolute protein content. Therefore, the two measurements may not always show a linear correlation.
Hello Naile Merve Güven
The MTT assay is used to measure cellular metabolic activity as an indicator of cell viability, proliferation, and cytotoxicity. The viable cells contain NADPH-dependent oxidoreductase enzymes which reduce the MTT to formazan. The insoluble formazan crystals are then dissolved using a solubilization solution and the resulting colored solution is quantified by measuring absorbance at 570nm. The darker the solution, the greater the number of viable, metabolically active cells.
On the other hand, the protein assay will measure the protein content of viable cells which is an indirect measurement of cell viability and evaluates cell toxicity though it is not a specific end point. The proliferation rate of the cells will decrease because of cytotoxicity and this in turn will bring about the reduction in protein content of the cultured cells.
Attached below are two papers where you will find cytotoxicity assays such as MTT, LDH and Neutral Red assay are compared with protein content.
Article Cytotoxic Effects of a Novel Thialo Benzene Derivative 2,4-D...
https://www.sciencedirect.com/science/article/abs/pii/S0378427405001967?via%3Dihub
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