A researcher is attempting to measure the protein concentration in an unknown sample using a standard curve generated from known concentrations of BSA (Bovine Serum Albumin). Which of the following factors is most critical for ensuring the accuracy of the protein quantification from the standard curve?

The most critical factor is the accurate preparation and measurement of the BSA standards that form the standard curve, because any error in those concentrations will propagate to every sample measurement. To keep the curve trustworthy, the standards must be made with precise dilutions, pipetted carefully, and assayed under exactly the same conditions (time, temperature, reagent volumes) as the unknown sample. If the standards are off, the curve’s slope and intercept will be wrong, and the calculated protein concentration will be systematically biased. For example, if a 0. 5 mg/mL BSA standard is mistakenly prepared as 0.