Quiz 3 key (2) - Biochemistry 309, Winter 2010 Quiz #3...

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Biochemistry 309, Winter 2010 20 points Quiz #3 NAME:_______________________________ TA:__________________ 1. After testing various conditions and purification steps in the process of purifying an enzyme, you determine the following: Your enzyme is NOT precipitated by 35% ammonium sulfate (but 50% of the total protein is). Your enzyme IS precipitated by 45% ammonium sulfate and is fully active when resuspended. Your enzyme binds to a cation exchange column at pH 7 in the presence of 25mM NaCl Your enzyme does not bind to the same cation exchange column either at pH 8 + 25mM NaCl or at pH 7 + 100mM NaCl. Your enzyme elutes earlier than a 50kDa protein from a gel filtration (size exclusion) column. What can you conclude about the approximate size of your enzyme? (2 points) The enzyme is >50KDa. What can you conclude about the approximate pI of your enzyme? (2 points) The pI of the protein is >7 and 8. Order the following purification steps in the best way to purify your enzyme.
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This note was uploaded on 02/16/2012 for the course BIOL_SCI 309 taught by Professor Somebody during the Winter '10 term at Northwestern.

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Quiz 3 key (2) - Biochemistry 309, Winter 2010 Quiz #3...

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