What kind of column are we using in class? a. Anton exchange chromatography. b.

Question

What kind of column are we using in class? a. Anton exchange chromatography. b. Antibody affinity chromatography. C. Ni-affinity chromatography. d. Liquid chromatography. In order to isolate a protein that is glycosylated, what kind of starting material can one use? a. A bacterial culture in which the protein of interest was produced (either constitutively or induced). b. A cell lsyate that you knew contained the protein of interest. c. A cellular extract from a eukaryotic gene expression system in which the protein of interest was produced. d. Two of the above e. All of the above (a-c). Density centrifugation is a technique that is used to isolate protein complexes. True False

Explanation / Answer

Q4 is based on the column which was in use in the class so cannot be answered.

Q5.All the options mentioned.

Glycosylation is an addition of carbohydrate moieties to the proteins once they are translated such that they are transported to specific locations.Glycosylation occurs both in prokaryotes and eukaryotes So one can use a bacterial culture, a cell lysate and a cellular extract can be used to obtain the protein of interest.In the case of bacterial culture if it is secreted outside it is very easy as the culture can be centrifuged and the protein can be isolated from the media.In cell lysate, one can perform chromatography where the protein binds to the column.

Q6. Density centrifugation is a process in which a solution containing various components could be carbohydrates proteins is taken and spun in specialised tubes at a specified gravity.This settles the densest component to settle down at the bottom of the tube and the lightest is on the top.This method will not give a pure compound but an easy initial screening procedure.

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