Question
Consider a crude extract with a mixture of the 5 proteins listed below. Protein A 4 . 5 pI 1 6 0 molecular weight, kDa
Consider a crude extract with a mixture of the proteins listed below.
Protein A pI molecular weight, kDa
Protein B pI and molecular weight
Protein C pI molecular weight
Protein D pI molecular weight
Protein E pI molecular weight
You load this protein mixture onto an anion exchange column at pH
Next, you apply a "washing" step by passing through buffer at pH
Finally, for your elution step, you apply a pH gradient starting from pH to pH A gradient buffer system allows you to gradually and continuously change the pH of your mobile phase starting from pH up to pH You load this same protein mixture onto a Size Exclusion column. Please indicate the order in which these proteins will elute for both. If your protein of interest is protein A would using anion exchange column be completely successful at separating it from all the other proteins? Please explain.
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Molecular Cell Biology
Authors: Harvey Lodish, Arnold Berk, Chris A. Kaiser, Monty Krieger, Anthony Bretscher, Hidde Ploegh, Angelika Amon, Matthew P. Scott
7th edition
1464183393, 1464183392, 978-1429234139
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