The following is the chromatogram of an anion exchange column. The absorbance, v
ID: 489648 • Letter: T
Question
The following is the chromatogram of an anion exchange column. The absorbance, volume of elution, and the salt gradient concentration are shown in the different axes. Two proteins, A and B were purified using the column. Answer the questions based on the chromatogram.
Charge of A:
Charge of B:
Which protein has more net charge to bind to the column tightly?
What salt concentration did A elute at?
What salt concentration did B elute at?
Which of the two proteins are bigger?
0.6 M 0.4 M 8 0.2M 15 25 35 55 65 75 85 Volume (mL) uonenueou00 IOeN MM B A 5 08ZVExplanation / Answer
CHARGE OF B IS + AS IT WASHED OUT AND CHARGE OF B IS -VE PROTEIN A HAS ANETCHARGE TO BIND TO THE COULUMN SALT CONCENTRATION FOR A IS 20 AND SALT CONCENTRATION OF B IS 50 AS PER SALT CONCENTRATION PROTEIN B IS BIGGER
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