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Exercise #1 prelab - Name_Wendy Mei 1(8 points Illustrate...

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Name_Wendy Mei________ 1. (8 points) Illustrate the structures of an amino group above and below its pKa of 8 and a carboxyl group above and below its pKa of 3.1. Amino Groups Carboxyl Group Above pKa of 8: NH 2 Above pKa of 3.1:COO - Below pKa of 8:NH 3 + Below pKa of 3.1: COOH 2. ( 2 pts) The pH at which a protein’s overall charge is zero is the __isoelectric point (pI)____. 3. (5 points) What charge must a protein have to stick to a DEAE column? We are using changes in salt concentration to elute the proteins. Why does this pull proteins off the column? A protein must have a negative charge to stick to a DEAE column because the DEAE groups are positively charged. The changes in salt concentration pulls proteins off the column because the chloride ions compete with the proteins to bind to the DEAE column. 4. (5 points)We will be separating three proteins, differing in pI, with buffers at two different pHs. Predict what you will see at each pH (i.e. how many peaks, what is the order).
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