Protein Purification
Protein Extraction
Differential Centrifugation
Salting Out
Column Chromatography
Ion-Exchange Chromatography
Anion-Exchange Chromatography
Size Exclusion Chromatography
Affinity Chromatography
Specific Activity
Native Gel Electrophoresis
SDS-PAGE Strategies
Isoelectric Focusing
Diagonal Electrophoresis
Mass Spectrometry
Mass Spectrum
Tandem Mass Spectrometry
Peptide Mass Fingerprinting
Overview of Direct Protein Sequencing
Amino Acid Hydrolysis
Chemical Cleavage of Bonds
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: Using Beta-Mercaptoethanol with SDS-PAGE

Practice: Compare the Native & SDS PAGE gels to indicate if each sample is a monomer, dimer, trimer or tetramer.

a. Sample 1: ________________

b. Sample 2: ________________

c. Sample 3: ________________

d. Sample 4: ________________

Practice: Protein X” has a molecular mass of 400 kDa when measured by size-exclusion chromatography. When subjected to SDS-PAGE, Protein X gives 3 bands with molecular masses of 180, 160, & 60 kDa. When SDS-PAGE is conducted a second time but in the presence of β-mercaptoethanol (β-ME), 3 bands form again, but this time with molecular masses of 160, 90, and 60 kDa. What is the subunit composition of Protein X? (Hint: draw both SDS-PAGE gels).