Pre-Lab: In addition to the procedure, prepare a dilution table for the BSA with total protein added to the assay, and calculate the total dilution needed for the ‘unknown’ milk sample Also, calculate the EXPECTED absorbance of a 1/5 dilution (300 μしtotal volume) of the 1.00 mg/mL BSA standard solution. Part A. BCA assay The BCA assay first requires dilutions of 1.0 mg/mL BSA from 0:100 to 100:0 in 10 μL increments in water. These dilutions can be prepared in the microplate, say row A or B, and shake by gently tapping the plate against the edge of the bench. be tor The unknown is a sample of non-fat milk which has a reported protein concentration of 34.0 mg/mL. This sample needs to be diluted such that the amount in 10 μL added to the BCA assay is within the range of the BSA amounts used for the standard curve (1.00-10.0 μg). The final dilution of the milk sample should be 10/290piL in BCA. Be sure that the milk BCA assays are set up at the same time as the BSA assays since results are time dependent. The dilute solutions of BSA (10 μL of the dilutions) can then be assayed with 290 μしof BCA reagent (tree green colored solution) in a VIS microplate. A protocol is set up -it reads C1-C11 for the standard curve, D1-D3 for the unknown sample, and displays the standard curve named “C485_BCA_Protein_Assay.” Otherwise, you’ll need to set up your own protocol. The BCA assay needs to be covered and incubated for at least 30 min in the incubator above 45°C. The absorbance is then read at 562 nm on the plate reader. Part B. Bradford assay of BSA The Bradford assay requires the same dilutions of BSA, and the assay is prepared with 10 μL ofdilute BSA and milk dilutions with 290 μL Bradford reagent in a VIS-BRADFORD plate. The plate is incubated at room temperature for at least 10 minutes and the absorbencies taken at 595 nm using the UV/VIS micro plate reader. A protocol is already setup, and reads the standard curve in cells C1-C11, the unknowns in D1- D3, and gives a standard curve, named C485 Bradford.