for the second step the starting concentration is 45%.
samita, on 19 March 2011 - 12:30 PM, said:
I want to precipitate the protein twice with amoinum sulfate. First 45 % and then 100 %. When I will do 45% initial amount of ammonium sulfate is 0. I will add the required am. sulfate and then centrifuge the solution and dicard the pellet and take the supernatent. This supertatent I want to precipitate to 100 %. For this step initail ceoncetration of am. sulfate is 0 or 45 % to count the amount of am. sulfate for second 100 % precipitation.