Question #2 a for a biochemistry lab question. I\'m confused as to what numbers
ID: 474280 • Letter: Q
Question
Question #2 a for a biochemistry lab question.I'm confused as to what numbers to use for the hydrolyzed protein and total protein in order to calculate the percent hydrolysis mple calculations are not required for question R1. Question The samples that were provided for analysis in thisexperiment were Casein (a protein found in milk) and Casein Hydrolysate (the protein broken down into the constituent amino acids). The per cent of Casein that was successfully hydrolyzed an be calculated from each assay. a) Biuret: hydrolysis. hydrolyzed protein /total protein x 100m. Remember that the Biuret assay quantifies intact protein therefore the amount ofhydrolyzed protein is obtained by calculating the inamount ofintactprotein before and after hydrolysis. Calculate the hydrolysis for test tubes n and 9 osmu aliquots were used and also do this calculation for testtubes 8 and 10 where 1mL aliquots were used. Show one sample calculation and then tabulate the data for both calculations. Lab coat, gloves and glasses must be worn. A waste from today'seperiment can be disposed of in the sink. 1. You wal need approximately smLeachofCasein, Casein Hydrolysate, and 10 melmu. Bovine Serum Albumin (BSAl (solutions were prepared inoosM phosphate buffer, pH 78). Put each solution in a separate 10 ml beaker and label (fia 10 mL beaker approximately half full to estimate 5 mL. 2. Bauretassar This assay will be used to determine the protein concentration of the Casein and Casein Hydrolysate solutions. The data from the 10 mg/mL BSA solution will be used to plot a calibration curve. Set up the assay as given below, using large (18x150) test tubes. Label your test tubes. You must pipet carefully because inaccurate volumes wil destroy your results. All the volumes in the table are in mL (1mL 1000wLJ. To each add 80 mL of Biuret reagent and mix thoroughly (vortex mixer) Alow this to stand for 30 minutes. Read 96 Tat550 nm against the reagent blank mube No. 11. Use only one cuvette to set 100 T (with the blank) and for all%Treadings. Save these samples until you have read all the NT values for the Biuret assay (discard the small amounts used for rinsing the cuvettes so that you do not dilute or contaminate your samples)
Explanation / Answer
First plot a calibration curve from the absorbance data of the tubes 2 to 6.
Be careful to use the correct concentrations (taking into account all the dilution factors).
Let's assume that the plot equation for the curve you obtained is:
A = k*C
Here k is the proportionality constant.
Now use this equation to find the C values for the test tubes 7 and 9 by putting the absorbace (A) values for these tubes into the equation.
Let's say you get C1 value for tube 7 and C2 value for tube 9.
C1 gives the intact protein content before hydrolysis while C2 gives the intact protein after hydrolysis ( because casein hydrolysate is formed by hydrolysis of casein ).
The difference between the C1 and C2 calue will give you the amount of protein that got hydrolyzed.