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: Isoelectric Focusing

Practice: At some point during isoelectric focusing, proteins stop moving through the gel because:

Practice: Electrophoretic separation at pH 6 of a sample mixture with Peptide #1 (MW 100) Peptide #2 (MW 200) and Peptide #3 (MW 400) would result in which of the following? (Note: the pI of each peptide occurs at pH 6).

Practice: Mark the approximate final position of the following tripeptide on the isoelectric focusing gel: Glu-Met-Asp.

Hint: calculate the isoelectric point of the peptide.