Bradford Assay Method

To determine the concentration of the ADH enzyme solution, you must prepare 2 protein standard curves, one with bovine gamma globulin (BGG), the other with bovine serum albumin (BSA).To prepare you standard curves, take 5 test tubes and label them 1 to 5

1. Prepare dilutions of the BGG solution in the tubes as indicated in Table 1.

2. Add 500μl Bradford Solution to each tube and mix carefully with the vortex mixer.

3. Incubate for 5 minutes

4. Transfer the solution to a 3ml cuvette

5. Set the wavelength of the photometer to 595nm, calibrate using Tube 1 mixture (see theinstructions on how to calibrate the spectrophotometer).

6. Take the absorbance at 595nm for all samples and record your data in Table 1

Question 1. Calculate the dilution factor and then the protein concentrations for each test tube

Question 2. Create a standard curve in excel absorbance @595nm against conc. of the standard proteins, (1 graph with 2 trendlines: 1 for BGG and 1 for BSA)

Question 3. Using the average absorbance of ADH (attached file with all the data) use the equations of the trendlines to determine the protein concentrations for each of the dilutions, calculate the average initial concentration of ADH according to the BGG and BSA curves.

Attachment:- Assay Method.rar