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5. 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 1 2 3 4 5 6 7 8 Protein Collagen y-Globulin Insulin Fibronectin Lysozyme Pepsin Serum Albumin Myoglobin Molecular Weight 300,000 150,000 5,700 220,000 14,300 34,500 66,500 16,700 pl Conc 0.5 mg/L 4.7 7.2 1 mg/L 12 mg/L 5.3 5 mg/L 5.8 2 mg/L 11.0 10 mg/L 1.0 4 mg/L 4.9 2 mg/L 7.1 - added prior to running IEF? -left out of the process entirely? Type fibrous a-helices globular protein globular protein fibrous protein enzyme digestive enzyme blood protein 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 flow down the page). Assume that the electrophoresis step is timed so that none of the proteins run off the end of the gel. Consider the different concentrations of each protein in the mixture when making the qualitative drawings. 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:

5. 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 1 2 3 4 5 6 7 8 Protein Collagen y-Globulin Insulin Fibronectin Lysozyme Pepsin Serum Albumin Myoglobin Molecular Weight 300,000 150,000 5,700 220,000 14,300 34,500 66,500 16,700 pl Conc 0.5 mg/L 4.7 7.2 1 mg/L 12 mg/L 5.3 5 mg/L 5.8 2 mg/L 11.0 10 mg/L 1.0 4 mg/L 4.9 2 mg/L 7.1 - added prior to running IEF? -left out of the process entirely? Type fibrous a-helices globular protein globular protein fibrous protein enzyme digestive enzyme blood protein 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 flow down the page). Assume that the electrophoresis step is timed so that none of the proteins run off the end of the gel. Consider the different concentrations of each protein in the mixture when making the qualitative drawings. 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:

Biochemistry
Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
Section: Chapter Questions
Problem 1P
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5. 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 1 2 3 4 5 6 7 8 Protein Collagen y-Globulin Insulin Fibronectin Lysozyme Pepsin Serum Albumin Myoglobin Molecular Weight 300,000 150,000 5,700 220,000 14,300 34,500 66,500 16,700 Conc pl 0.5 mg/L 4.7 1 mg/L 7.2 12 mg/L 5.3 5.8 11.0 5 mg/L 2 mg/L 10 mg/L - added prior to running IEF? left out of the process entirely? 4 mg/L 2 mg/L 1.0 4.9 7.1 A. Which of these proteins would electrophoresis NOT separate effectively? B. Which of these proteins would isoelectric focusing NOT separate effectively? Type fibrous a-helices globular protein globular protein fibrous protein enzyme digestive enzyme blood protein muscle protein 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 flow down the page). Assume that the electrophoresis step is timed so that none of the proteins run off the end of the gel. Consider the different concentrations of each protein in the mixture when making the qualitative drawings. 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:
Transcribed Image Text:5. 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 1 2 3 4 5 6 7 8 Protein Collagen y-Globulin Insulin Fibronectin Lysozyme Pepsin Serum Albumin Myoglobin Molecular Weight 300,000 150,000 5,700 220,000 14,300 34,500 66,500 16,700 Conc pl 0.5 mg/L 4.7 1 mg/L 7.2 12 mg/L 5.3 5.8 11.0 5 mg/L 2 mg/L 10 mg/L - added prior to running IEF? left out of the process entirely? 4 mg/L 2 mg/L 1.0 4.9 7.1 A. Which of these proteins would electrophoresis NOT separate effectively? B. Which of these proteins would isoelectric focusing NOT separate effectively? Type fibrous a-helices globular protein globular protein fibrous protein enzyme digestive enzyme blood protein muscle protein 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 flow down the page). Assume that the electrophoresis step is timed so that none of the proteins run off the end of the gel. Consider the different concentrations of each protein in the mixture when making the qualitative drawings. 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:
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