I will measure absorbance for measurement of protein concentration.
The sample solutions are prepare in serial dilutions to the volume of 100 uL to obtain a certain concentration of the protein in the buffer, lets say 2 mg/mL, then 20 uL is added to the cuvette with 1 mL Bradford reagent.
Now my question is, is the protein concentration still 2 mg/mL, I mean, I know it is not in the cuvette with the reagent added, but do I have to take this into account when measuring the absorbance? Or should I recalculate the dilutions to get the concentration 2 mg/mL in total with the reagent added?
This might be a dumb question but the hour is late and my brain is scrambled eggs.
Thank you in advance