1 / 8

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.

dyllis
Download Presentation

Biochemistry Lab

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript


  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

More Related