Clutch Prep is now a part of Pearson
Ch. 5 - Protein TechniquesWorksheetSee all chapters
All Chapters
Ch. 1 - Introduction to Biochemistry
Ch. 2 - Water
Ch. 3 - Amino Acids
Ch. 4 - Protein Structure
Ch. 5 - Protein Techniques
Ch. 6 - Enzymes and Enzyme Kinetics
Ch. 7 - Enzyme Inhibition and Regulation
Ch. 8 - Protein Function
Ch. 9 - Carbohydrates
Ch. 10 - Lipids
Ch. 11 - Biological Membranes and Transport
Ch. 12 - Biosignaling
Clutch Review 1: Nucleic Acids, Lipids, & Membranes
Clutch Review 2: Biosignaling, Glycolysis, Gluconeogenesis, & PP-Pathway
Clutch Review 3: Pyruvate & Fatty Acid Oxidation, Citric Acid Cycle, & Glycogen Metabolism
Clutch Review 4: Amino Acid Oxidation, Oxidative Phosphorylation, & Photophosphorylation
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: Anion-Exchange Chromatography

Example #1: Circle the peptide below that elutes last during anion-exchange chromatography if the pH is ~7?

Practice: Which amino acid elutes last from an anion-exchange column at physiological pH?

Practice: Use the chart to determine which tripeptide would elute last from an anion-exchange column at pH = 9.3.

Practice: Which type of ion exchange chromatography would be best to separate a mixture of histidine and arginine? 

            His: pK1 = 1.8, pK2 = 9.3, pKR = 6.0                     Arg: pK1 = 1.8, pK2 = 9.0, pKR = 12.5

Practice: Stationary resin compounds with carboxymethyl (CM) and diethylaminoethyl (DEAE) groups are shown below. Indicate which one is likely used in a cation exchange column and which one is likely used in an anion exchange column. Considering the following peptide at pH 7, should DEAE or CM groups be used as the stationary resin to purify the peptide? 

Peptide: G-R-W-K-R-H