Step Sample Total rGFP activity (RFU) Bradford measred total protein (ug/ml) SDS-page measures purity % A GCE(1ml) 48,000 450 5
B Pool E fraction from Ni^+2 agarose (1ml) 35,000 98 40
C Prooled F fraction from anion exchanger 30,000 9 85
D Pooled GF fractions from Sephadex G-100 29,000 5 90

Using the protocol and the purification table above to answer the following questions:

a. Going from what two steps resulted in the largest increase in purity?
b. Going from what two steps resulted in the lowest loss of activity?
c. How many ug of total protein was applied to the gel filtration column? Show your calculations for credit?
d. How much rGFP in ug was present at the beginning of the purification process? Show your calculations for credit (2018)?
e. Going from what two steps resulted in the greatest fold decrease in total protein? Show your work for credit (2008)?
f. What was the yield in g of rGFP finally purified using this purification process? Show your calculations for credit.
g. In an attempt to save money and time, which step would step would be the best step to remove without significantly altering the purity of rGFP?