Protein Purification Manual
Purification of ß-galactosidase from E. Coli
Jeanette Mowery and Lisa Seidman
Biotechnology Lab Technician Program
Madison Area Technical College
Funded in part by NSF ATE grants
DUE 9752027
The printable version of this document.
Table of Contents
Chapter 1: Introduction
Introduction
Chapter 2: Protein Structure
Protein Structure
Chapter 3: Assays and Calculations
Assays and Calculations
Procedures:
3.1 Bradford Microassay
Sample Calculation for Bradford Assay
3.2 ß-Galactosidase Assay
Sample Calculation for ß-Galactosidase Assay
Sample Calculation for Specific Activity
Sample Calculation for Purification Factor
Sample Calculation for Yield
Sample Calculation for Percent Yield
Summarize Purification Results
Chapter 4: Purification Methods
Purification Methods
4.1 Making the Crude Extract
4.2 Ammonium Sulfate Precipitation
4.3 Dialysis: Desalting and Buffer Exchange
4.4 Ion Exchange Chromatography
Results and Data Analysis
Chapter 5: Analytical and Verification
Analytical and Verification
5.1 Concentration of Samples
5.2 Polyacrylamide Gel Electrophoresis
5.3 Staining the Gel
5.4 Western Blotting
Chapter 6: Reports
Reports
Appendix A: Solutions
Solutions
Appendix B: Materials and Order Information
Materials and Order Information
Appendix C: Resources
Resources
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