Question 20: B) & C)
(B) The purification protocol included application of 50mMNa,Na-buffer, pH 7.4 containing 10mM NaCl, 90mM KCL, 0.1% Tween-80,and 0.01% sodium azide. You have found following stock solutions inthe lab previously prepared by a reliable lab member: a) 1MNa,Na,-phosphate buffer, pH 7.4 b) 2M NaCl c) 1M KCl d) 10% Tween80 A small bottle of crystalline NaN3 was provided by your researchsupervisor. Calculate volumes (ml or µl) of each (a) through (d)stock solutions and the amount (mg or µg) of NaN3 need to be mixedtogether in order to prepare 250ml of required buffer.
(C) A purified protein is a metalloprotein which has amononuclear zinc binding site. The purification procedure includedseveral harsh conditions causing the protein to lose its metal,thus converting it into its apo-form. Calculate the amount of a20mM ZnCl2 solution must be added to a 2ml aliquot of the proteinin order to get a fully metallated form for furtherinvestigation.
