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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 5 proteins listed below.

Protein A 4.5 pI 160 molecular weight, kDa

Protein 12.5 pI and 65 molecular weight

Protein C 5.0 pI, 15 molecular weight

Protein D 6.8 pI, 150 molecular weight

Protein E 9.5 pI, 45 molecular weight

You load this protein mixture onto an anion exchange column at pH 11.

Next, you apply a "washing" step by passing through buffer at pH 11.

Finally, for your elution step, you apply a pH gradient starting from pH 11 to pH 2.0. (A gradient buffer system allows you to gradually and continuously change the pH of your mobile phase starting from pH 11 up to pH 2). 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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