You start with 100 units of protein activity and 100 grams of total protein. After the first centrifugation step, you have 25% less activity and 85% less total protein. What is your fold-purification of the target protein? 0.3-fold 3.4-fold 75-fold 5-fold 20-fold

4) Researcher #1 and #2 in your lab each conducted a protein purification. They present you with the following data. The protein-of-interest is indicated by the red arrowhead, and the numbers on top indicate the purification step. Researcher #1 1 2 3 4 SDS-PAGE (Coomassie Stained) Researcher #2 1 2 3 4 5 6 Immunoblot a) Which researcher has more clearly demonstrated purification of the protein-of-interest? Justify your answer. b) What type of chromatography step did researcher #1 most likely conduct in going from step 2➜ 3 based on the gel? Justify your answer.

Explain the methods of Protein Engineering?  Please answer at your own words.

After properly troubleshooting, you finally are able to visualize the gel. You obtained the image below. The ladder used is “Lambda Hind III”.Knowing that the ladder “Lambda Hind III” consists of six fragments (23130 bp, 9416 bp, 6557 bp, 4361 bp, 2322 bp, 2027 bp), properly label the bands in the gel image.

In DNA purification, explain how the chromosomal DNA is separated from the plasmid DNA? Be sure to mention the specific buffer components that facilitate this process.

Protein analysis by gel electrophoresis  a).  Using the gel image provided to calculate the electrophoretic relative mobility as a ratio of the distance of protein migration to the distance of the tracking dye migration (See Appendix B).  If your dye front is not visible, measure the mobility relative to the bottom edge of the resolving gel.  Include a labeled print-out of your gel image with your report.  b).   Plot log MW of the protein markers and commercial myoglobin vs. mobility. Determine the molecular weight of myoglobin obtained after the final purification step (Sample E) from the equation of the line.  Submit a copy of your graph along with the gel image. c). Compare your sample E and commercial myoglobin with the ladder. Summary your results.   order of sample (left to right) is :  A  B  C  D  blank  E  blank  Commercial Myoglobin   Molecular Ladder