Bradford Assay to determine Protein Concentration

Materials:

 * Bradford Assay Kit (Amresco)
 * Protein Sample (concentration to be determined)

Method:

 * 1) Into 6 separate microcentrifuge tubes, aliquot 5, 10, 15 and 20 µL of 0.5 mg/ml BSA solution. Bring the volume of each to 100 uL with desired buffer.
 * 2) Label a tube as BLANK and pipette 100 uL of buffer into it. Use this tube to blank the spectrophotometer.
 * 3)    Add to each tube, 1 mL Bradford Reagent and vortex. Allow to stand at room temperature for 2 minutes.
 * 4)    Determine the absorbancies at A595 nm using a 1 mL microcuvette.
 * 5) Generate a standard curve by plotting absorbance at 595 nm versus protein concentration.
 * 6) For your protein sample, repeat the same steps using your protein sample in place of the BSA. Plot the A595 nm and use the standard curve as a reference to determine the concentration of the unknown protein sample.