Introduction To determine the purity of a protein, a sodium dodecyl sulfate polyacrylamide gel …show more content…
Then centrifugation is used to remove all the non-protein macromolecules, resulting in a large number of bands in the CCE lane. Most of the bands in the lane are very faint because they are not concentrated in the whole protein sample. In the Flow-Through stage, MBP-tagged AP binds to the amylose resin and the rest is eluted off the column, as reflected in the FT lane. In this stage, most of the undesired protein is removed. For Wash 1, Wash 2, and Wash 3, every last bit of undesired protein is eluted of the column. However, in Wash 2, there is a small amount of the MBP-AP protein that was eluted off the column, which mean be caused by human errors such as cross contamination. Finally, in Elution 1 and Elution 2, elution buffer was used to elute MBP-AP off the column, resulting in a strong band in E1 and E2 lanes. In addition, there are no visible impurities, which means that the protein is very