Table of contents

1. Introduction to Biochemistry4h 34m
2. Water3h 23m
3. Amino Acids8h 10m
4. Protein Structure10h 4m
5. Protein Techniques14h 5m
6. Enzymes and Enzyme Kinetics13h 38m
7. Enzyme Inhibition and Regulation 8h 42m
8. Protein Function 9h 41m
9. Carbohydrates7h 49m
10. Lipids5h 49m
11. Biological Membranes and Transport 6h 37m
12. Biosignaling9h 45m
Review 1: Nucleic Acids, Lipids, & Membranes2h 47m
Review 2: Biosignaling, Glycolysis, Gluconeogenesis, & PP-Pathway3h 12m
Review 3: Pyruvate & Fatty Acid Oxidation, Citric Acid Cycle, & Glycogen Metabolism2h 26m
Review 4: Amino Acid Oxidation, Oxidative Phosphorylation, & Photophosphorylation1h 48m

5. Protein Techniques

Anion-Exchange Chromatography

Biochemistry

5. Protein Techniques

Anion-Exchange Chromatography

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Multiple Choice

Which type of ion exchange chromatography would be best to separate a mixture of histidine and arginine?
His: pK₁ = 1.8, pK₂ = 9.3, pK_R = 6.0 Arg: pK₁ = 1.8, pK₂ = 9.0, pK_R = 12.5

Anion-exchange chromatography at pH = 2.

Anion-exchange chromatography at pH = 4.

Cation-exchange chromatography at pH = 2.

Cation-exchange chromatography at pH = 4.

Cation-exchange chromatography at pH = 9.

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Verified step by step guidance

Understand the principle of ion exchange chromatography: It separates molecules based on their net charge at a given pH. Cation-exchange chromatography uses a negatively charged resin to bind positively charged molecules, while anion-exchange chromatography uses a positively charged resin to bind negatively charged molecules.

Determine the net charge of histidine and arginine at different pH levels: Use the pKa values provided to calculate the net charge of each amino acid at the specified pH levels. The net charge is determined by the ionization state of the amino acid's functional groups.

Calculate the isoelectric point (pI) for each amino acid: The pI is the pH at which the amino acid has no net charge. For histidine, the pI is calculated using the average of pK2 and pKR, and for arginine, it is the average of pK2 and pKR.

Evaluate the net charge of histidine and arginine at pH 9: At this pH, compare the net charges of histidine and arginine to determine which will be positively charged and thus bind to a cation-exchange resin.

Select the appropriate ion exchange method: Since both histidine and arginine will have different net charges at pH 9, cation-exchange chromatography is suitable for separating them based on their positive charges.