The purpose of this experiment was to determine the concentration of Bovine Serum Albumin protein through the use of the Lowry Assay. This was completed by first creating a standard curve from known concentrations of the Bovine Serum Albumin protein. First, it was essential to create ‘blank’ using water, and to measure the absorbance of the blank through the spectrophotometer. We were able to create several standards with known concentrations of BSA that included both low and high concentrations. After running the assay, we were able to graph a standard curve by plotting the known protein concentrations, in mg/L, against the spectrophotometer readings of absorbance. From this standard curve, the concentration for an unknown can be determined using its corresponding reading of absorbance.
It is important to mention that one point was removed from the data set when creating the standard curve graph that would give the equation of the line. One particular data point, which was a concentration of
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These color compounds were a result of the reaction that takes please in the Lowry Assay. In an alkali solution, the samples of protein are put into contact with copper. This results in the aromatic residues of both Tyrosine and Tryptophan reducing the Folin phenol reagent. This results in the previously treated sample’s aromatic amino acids reducing the Folin phenol reagent. In addition, the Folin phenol reagent is also treated with copper. This reaction results in a product with a blue color, and in order to determine the protein amount in each sample, it is necessary to read its absorbance at 660nm and compare that against a standard curve of Bovine Serum Albumin. It is essential to incorporate the color product of the reaction that reduces the Folin phenol reagent, as the spectrophotometer does not have the ability to detect and uncolored compounds