Objective
Make calibration curves using the Bradford Assay.
Procedure
- Make a stock solution that is roughly 10mg protein in 2mL of buffer.
- Calculate your actual concentration
- Make 6 standard solutions (1mL each) between 1μg/mL and 10μg/mL
- Determine the appropriate volume of your stock to use (for the proper final concentration in 1mL) and add that volume to an eppendorf tube
- Add 200μL of the Bio-Rad Protein Assay reagent
- Add the correct amount of buffer such that the final volume is 1mL
- Take a UV-Vis (no less than 1 hour after they were produced).
- Make 2 blanks as well (800uL buffer and 200uL Assay reagent) and take it's UV spectrum. (400nm-800nm)
- After you have finished one set, repeat the process (make new samples and new measurements)
Make Atomic Absorption standards for tomorrow!
Using the gold AA/ICPMS standard solution make 5 new solutions (Note: Use water - NOT BUFFER - to make these solutions)
- 25 ug/mL
- 20 ug/mL
- 15 ug/mL
- 10 ug/mL
- 5 ug/mL
Data (Trial 1)
- Series 1: 5 ug/mL
- Series 2: 6 ug/mL
- Series 3: 7 ug/mL
- Series 4: 8 ug/mL
- Series 5: 9 ug/mL
- Series 6: 10 ug/mL
Data (Trial 2)
- Series 1: 5 ug/mL
- Series 2: 6 ug/mL
- Series 3: 7 ug/mL
- Series 4: 8 ug/mL
- Series 5: 9 ug/mL
- Series 6: 10 ug/mL
Notes
- All data was collected using the shortened path length of 0.412 cm.
- Stock solution: 4950 ug/mL
- Trial 1: 0.0099 g protein
- Trial 2: 0.012 g protein
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