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: Protein Activity

Concept #2: Protein Specific Activity

Practice: A biochemist discovers and purifies a new enzyme and generates the purification table below.

A) Fill-in the table below with the specific activity of the enzyme after each purification procedure.

B) According to the data, which purification step was most effective to give the greatest relative increase in purity? ____________________________

C) Which purification step was least effective to give the lowest relative increase in purity? ____________________________

D) Based on the results in the table, should the biochemist be convinced that the enzyme is pure after step 6? Why?