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- Hello, I would like help with problem 42 in the image attached.-858kcal to mcalCould someone explain what this is saying? I would like a better understanding. FIGURE D CPP32 (Caspase-3) protease activity. Aliquots of cell lysates (50μL) were incubated with an equal volume (50 μL) of the reactionbuffer and 5 μL of 7-amino-4-trifluoromethyl coumarin (AFCDEVD) for 1 hour at 37˚C (Clontech). The shift in fluorescenceemission of AFC-DEVD, on its proteolysis to free AFC by theprotease, was detected in a fluorometer, using a 400 nm excitationfilter and 505 nm emission filter.