Why must you only add the enzyme after all the other reagents have been added?  Because until all reagents have been added the reaction mixture will not have the correct pH and/or ionic strength which may result in the enzyme been denatured and/or having reduced activity. Because until all reagents have been added the reaction mixture will not be at the correct temperature which may result in the enzyme been denatured and/or having reduced activity. Because until all reagents have been added the reaction mixture will not have the correct amount of substrate which may result in the enzyme not working optimally. Because the enzyme needs to be kept cold as long as possible, so it is not denatured and/or having reduced activity.

Biochemistry
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ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
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Chapter1: Biochemistry: An Evolving Science
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Why must you only add the enzyme after all the other reagents have been added? 

Because until all reagents have been added the reaction mixture will not have the correct pH and/or ionic strength which may result in the enzyme been denatured and/or having reduced activity.

Because until all reagents have been added the reaction mixture will not be at the correct temperature which may result in the enzyme been denatured and/or having reduced activity.

Because until all reagents have been added the reaction mixture will not have the correct amount of substrate which may result in the enzyme not working optimally.

Because the enzyme needs to be kept cold as long as possible, so it is not denatured and/or having reduced activity.

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