Assessment of Total Cellular Protein Following Fatty Acid Treatment Using the BCA Assay

Carli B. Maguire

Fatty acids can influence cellular metabolism and protein homeostasis, potentially altering total protein content within cell samples. Accurate quantification of total protein concentration is essential for standardization for further biochemical assays and ensuring reproducibility. In this study, adherent mammalian cells were treated with a defined fatty acid preparation under controlled culture conditions. Following treatment, whole-cell lysates were collected and total protein concentration was determined using the bicinchoninic acid (BCA) assay. The BCA assay is a colorimetric method based on the reduction of Cu²⁺ to Cu¹⁺ by peptide bonds under basic conditions, followed by formation of a purple-colored complex between bicinchoninic acid and Cu¹⁺. Absorbance was measured at 562 nm and protein concentrations were calculated using a bovine serum albumin (BSA) standard curve. Preliminary findings demonstrate significant statistical differences in total protein concentration between treated and control groups, highlighting the importance of protein standardization when evaluating treatment-induced cellular responses. These results emphasize the utility of the BCA assay as a reliable method for quantifying total protein in cultured cells exposed to metabolic modulators.