Biol. 6250
Protein Purification Techniques
Spring 2006
Textbook: Protein Purification: Principles and Practice
Third Edition, ROBERT K. SCOPES
|
Date |
Topic |
Chapter |
|
1/10, 1/12 |
Introduction: Review of Protein Structure; Protein Assays |
3.1 |
|
1/17, 1/19 |
Enzyme Assays; Bookkeeping; Centrifugation |
3.2-3.5; 1.2 |
|
1/24, 1/26 |
Extracting Cells; Isolating Organelles; Stabilizing Proteins; Concentration and Desalting Protein Solutions |
2.1-2.4; 12.2; 1.4; 8.4 |
|
1/31, 2/2 |
Purification Methods based on Protein Solubility; "Salting Out", Alcohol, Isoelectric and Heat Precipitation |
4.1-4.7 |
|
2/7, 2/9 |
Theory of Liquid Chromatography |
5.1-5.6 |
|
2/14, 2/16, 2/21 |
Types of Chromatography: Ion Exchange Chromatography, Gel Permeation Chromatography, Adsorption and Reverse Phase Chromatography |
6.1-6.2; 8.1; 6.3-6.5 |
|
2/23 |
Practical Techniques in DEAE and Gel Permeation Chromatography |
6.2; 8.1 |
|
2/28 |
Chromatography Materials used as "Batch Methods" |
|
|
3/2 |
FIRST EXAM |
5.3 |
|
3/7 |
Density Gradient Ultracentrifugation of Proteins |
Handout |
|
3/9 |
Electrophoresis of Proteins |
8.2 |
|
3/21 |
Non-denaturing Electrophoresis |
11.1 |
|
3/23 |
SDS-PAGE; Isoelectric Focusing |
8.2; 11.1 |
|
3/28 |
Analysis of Electrophoresis Gels; Staining and Densitometry |
11.1 |
|
3/30 |
Immunoblotting or "Western Blotting" |
11.1 |
|
4/4 |
Criteria of Purity for Isolated Proteins |
11.1-11.2 |
|
4/6, 4/11 |
Chemical and Physical Characterization of Pure Proteins |
Handout |
|
4/13 |
Theory of Affinity Chromatography |
7.1-7.5 |
|
4/18 |
Affinity Chromatography-continued |
|
|
4/20 |
Recombinant DNA Methods: Fusion Proteins; Antibodies |
9.1-9.3 |
|
4/27 |
FINAL EXAM |
11:00-1:30 Thursday |
REMINDERS:
Dr. Putnam-Evans' office is BN-209; e-mail: evansc@mail.ecu.edu; phone 328-4811.
Dr. Hager’s office is BS-404; email: hagerp@mail.ecu.edu; phone 328-1848.