5. (10 pts) The proteins listed below are separated by (1) isoelectric focusing (IEF) followed by (2) sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). A single sample of the protein mixture is placed at the top center of the gel to begin step 1. ID Protein Molecular Weight Conc pl Type 1 Collagen 300,000 0.5 mg/L 4.7 fibrous a-helices 2 V-Globulin 150,000 1 mg/L 7.2 globular protein 3 Insulin 5,700 12 mg/L 5.3 globular protein 4 Fibronectin 220,000 5 mg/L 5.8 fibrous protein 5 Lysozyme 14,300 2 mg/L 11.0 enzyme 6 Pepsin 34,500 10 mg/L 1.0 digestive enzyme 7 Serum Albumin 66,500 4 mg/L 4.9 blood protein 8 Myoglobin 16,700 2 mg/L 7.1 muscle protein A. Which of these proteins would electrophoresis NOT separate effectively? B. Which of these proteins would isoelectric focusing NOT separate effectively? C. Show the expected results after each step, by drawing qualitative diagrams similar to those shown in the class notes. (consider the IEF to be left-right, with the anode located on the left to attract the low pl proteins and electrophoresis to go down the page). Assume that the electrophoresis step is timed so that none of the proteins run off the end of the gel. D. Qualitatively, what would happen to this process if the polyacrylamide gel were crosslinked more, meaning that the pores available for protein diffusion in the gel were smaller? E. What would happen if the sodium dodecyl sulfate (SDS) were: - added prior to running IEF? - left out of the process entirely