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You have a 25kD histidine tagged bacterial protein (100% pure).You apply this pure protein to a Ni2+-agarose column. Then you apply crude mouse cell extract to the column. After extensivewashing, you apply an imidizole buffer. Analysis of the resultantelution on an SDS-PAGE gel indicates two proteins (25kD and 74kD)of equalcoomassie blue staining.

1) What is the simplest explanation for why the 25kD and the75kDpurified off the column?

2) What would be an alternative explanation?

3) What control experiment could you perform to distinguish between the two possible conclusions you stated above?

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