You are given the following SDSPAGE gel to interpret Lane 1

You are given the following SDS-PAGE gel to interpret. Lane 1 contains molecular weight markers. The gel on the right (from the manufacturers website) is of the same standard MWM mixture, and provides the molecular weights for each protein. Do you think Coomassie Blue or Western Blot staining was used to visualize this gel? Briefly, explain your answer. Why might the molecular weight markers migrate differently (i.e.: have different spacing) in the two different gels? Which protein is smaller, A or E? Briefly, explain your answer. Approximately what molecular weight is protein B? You have the mixture of proteins that was loaded in lane 2 of the gel in question 2. Using the information provided above, design a good column chromatography procedure using the tools learned in class that will separate out protein B from the mixture. You will need to describe the column that you selected, any buffers required, and any steps required to recover the protein.

Solution

2a) Coomasie blue is generally used dye in SDS PAGE. The sulphonic acid group of the dye binds with amine group of proteins and gives blue color.

2b . Molecules with less molecular weight can move faster and cover more distance in less time as they can pass through the pores of gel easily . While molecules with higher molecular weight move slowly thus covering less distance .

c) E is the smallest molecule as it has travelled farthest distance in a given time. Since it is small so it can travel through pores eadily thus travel faster.

d )Molecular weight of B is around 25 kD