This City College Biochemistry lab report details the use of spectrophotometry at 595 nm to determine protein concentrations. Six known samples and two unknowns were analyzed, with a standard curve generated using BSA as a reference. This report provides clear data and insights into protein quantif...
Protein assays are vital techniques for quantifying protein concentration in biological samples.
One common method involves measuring the absorption of UV light by aromatic amino acids
within proteins, though this method has certain limitations. In this experiment, we determined
protein concentration using spectrophotometry at a wavelength of 595 nm. Six prepared samples
were measured for absorbance, along with two unknown samples.To quantify the unknown
protein concentrations, a standard curve was generated using BSA as a reference. Absorbance
and concentration exhibit a linear relationship, which allowed for the creation of a linear
standard curve. By plotting absorbance against known concentrations and applying the linear
equation (y = mx + b), we determined that unknown A, with an absorbance of 0.0475, had a
concentration of 0.060 mg/mL, and unknown B, with an absorbance of 0.082, had a
concentration of 0.086 mg/mL. The final concentrations included a 25x dilution factor.
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