This is a BIochem Lab Question Help Please You are given the

This is a BIochem Lab Question. Help Please!

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 (ie: 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

Coomassie blue dyes are a family of dyes commonly used to stain proteins in SDS-PAGE gels. Western blot staining is done on blots (nitrocellulose or PVDF membranes after the transfer of proteins from gel to the membranes)

Rate of Protein migration differs as the concentration of acrylamide in the gel varies. At low concentration of acrylamide the gels are more porous hence proteins migrate much faster while in high concentration acrylamide gels proteins migrate slow and separate better.

Proein mobility on gel depends on their molecular weight. As proteins move from negative electode to positive electrode the high molecular proteins drag and move slower than the low molecular weight protiens. A is high mol weight compared to E