Question: Chromatography have a limited protein binding capacity. During the Ni+2 Agarose Column lab, you noticed that the wash 2 (W2) fluoresced slightly. Based on the Western Blot antibody procedure, would you expect to see a band in the W2 lane? If sp wjay anpit be its molecular weight? What does this expected data day about the physical protein structure of rGFP?

Note: the Wash 2 was not overloaded in the Ni+2 agarose column. The protein used is rGFP, which contains His 6 tag on its
N-terminus and express epitope tag on its C-terminus. The MW of the enitre rGFP protein is 33 kD and His 6 tag and express epitope is 6 kD each. Not that the antibody binds at the express epitope tag and not the His 6 tag.