Sections
Protein Purification
Protein Extraction
Differential Centrifugation
Salting Out
Dialysis
Column Chromatography
Ion-Exchange Chromatography
Anion-Exchange Chromatography
Size Exclusion Chromatography
Affinity Chromatography
Specific Activity
HPLC
Spectrophotometry
Native Gel Electrophoresis
SDS-PAGE
SDS-PAGE Strategies
Isoelectric Focusing
2D-Electrophoresis
Diagonal Electrophoresis
Mass Spectrometry
Mass Spectrum
Tandem Mass Spectrometry
Peptide Mass Fingerprinting
Overview of Direct Protein Sequencing
Amino Acid Hydrolysis
FDNB
Chemical Cleavage of Bonds
Peptidases
Edman Degradation
Edman Degradation Sequenator and Sequencing Data Analysis
Edman Degradation Reaction Efficiency
Ordering Cleaved Fragments
Strategy for Ordering Cleaved Fragments
Indirect Protein Sequencing Via Geneomic Analyses

Concept #1: Affinity Chromatography

Practice: In your own words, describe the principles involved in protein purification by affinity chromatography.

Practice: The target protein to be purified is likely eluted from the affinity chromatography column by _______________. Explain potential advantages & disadvantages of the elution strategies.

Practice: A biochemist is attempting to separate a DNA-binding protein (protein X) from other proteins in solution (proteins A, B & C). Consider the chart & answer the questions below about what type of technique is best for separation.

A. What type of chromatography is best for separating protein X from protein A? ___________________________

B. What type of chromatography is best for separating protein X from protein B? ___________________________

C. What type of chromatography is best for separating protein X from protein C? __________________________