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Biochemistry Lab

Biochemistry Lab. Protein Purification - 2 Chromatography. Methods of Cell Lysis. Passive Methods: Freeze-thaw Chemical Enzymatic Osmotic Pressure. Physical Methods: Sonication Cell Bomb Homogenizer. Additional Considerations:. Volume Cell Type Toxicity Cost.

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Biochemistry Lab

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  1. Biochemistry Lab Protein Purification - 2 Chromatography

  2. Methods of Cell Lysis • Passive Methods: • Freeze-thaw • Chemical • Enzymatic • Osmotic Pressure • Physical Methods: • Sonication • Cell Bomb • Homogenizer Additional Considerations: • Volume • Cell Type • Toxicity • Cost • Further Purification Steps • Protein Stability • Protease activity/inhibitors • Protein location • pH • Salt • Reducing Agents

  3. Chromatography

  4. Types of Chromatography

  5. Gel Filtration

  6. FPLC

  7. FPLC – not so much….

  8. In lab this week PectateLyase Number of amino acids: 344 Molecular weight: 36752.1 Theoretical pI: 8.20 • Pour a 12% SDS-PAGE gel • Use gravity flow Ion Exchange chromatography to purify pectatelyase

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