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

Protein Purification

Biochemistry

5. Protein Techniques

Protein Purification

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Multiple Choice

A scientist is looking to study a specific protein called mitochondrial transcription factor A (TFAM). Which of the following is the most appropriate sequence of steps for protein purification of TFAM?

Extraction → Dialysis → Salting Out → Differential Centrifugation → Chromatography.

Extraction → Chromatography → Differential Centrifugation → Salting Out → Dialysis.

Extraction → Differential Centrifugation → Salting Out → Dialysis → Chromatography.

Extraction → Salting Out → Dialysis → Chromatography → Differential Centrifugation.

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Verified step by step guidance

Begin with the extraction step to isolate the protein from the cellular components. This involves breaking open the cells to release their contents, including the protein of interest, TFAM.

Use differential centrifugation to separate the cellular components based on their size and density. This step helps in removing larger debris and organelles, enriching the sample for the protein of interest.

Apply the salting out technique, which involves adding a high concentration of salt to the solution to precipitate the protein. This step helps in concentrating the protein and removing some impurities.

Perform dialysis to remove the excess salt and small molecules from the protein solution. This step is crucial for preparing the protein for further purification steps.

Finally, use chromatography to achieve high purity of the protein. This technique separates proteins based on their size, charge, or affinity, allowing for the isolation of TFAM in its pure form.