Consider a crude extract with a mixture of the 5 proteins listed below. Protein A 4 .
Question:
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.
Molecular Cell Biology
ISBN: 978-1429234139
7th edition
Authors: Harvey Lodish, Arnold Berk, Chris A. Kaiser, Monty Krieger, Anthony Bretscher, Hidde Ploegh, Angelika Amon, Matthew P. Scott