Protein isolation

biotechnology presentation

1. PROTEIN ISOLATION

2. PRESENTATION BY MARRIYAM BATOOL ANITA BANO AQSA ABBASI SANA HABIB

3. WHAT ARE PROTEINS? Proteins are the bio-molecules composed of amino acids; forming the building block of the system and performs most of the biological functions of the system.

4. WHAT IS PROTEIN ISOLATION? The process by which the proteins from the cells or tissues are recovered for the analysis purpose is known as protein isolation or protein extraction.

5. HISTORY proteins were known as the distinct class of biological molecules They distinguished this by its ability to coagulate Studies focused on the proteins that could be purified in large quantities

6. WHY WE PERFORM PROTEIN EXTRACTION? To compare the structure of proteins b y different organism To purify a protein in order to identify the gene that encodes it and to resolve proteins by SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis)

7. To examine an enzyme in a crude extract for physiological studies To study the mechanism of action of an enzyme To diagnose parasitic disease

8. METHODS FOR PROTEIN ISOLATION

9. Chromatography Ultrasound Homogenisation French Press Cryogenic Grinding (Grinding in Liquid Nitrogen) Lysis Buffer

10. CHROMATOGRAPHY Ion Exchange Chromatography Separates proteins based on their surface ionic charge Proteins with very similar charges can be separated Ion exchange column utilized the principle of ion exchange chromatography

11. 2. Gel Filtration Chromatography Also known as size-exclusion or gel-permeation chromatography Separates proteins according to size and shape Separation of molecules is divided into three main types

12. ULTRASONIC HOMOGENISATION Used for tissues like some leaves and a post treatment after grinding Does not require freezing thus may avoid artefacts of freezing but may cause artefacts by heating of the sample

13. FRENCH PRESS Used for individual cells (plant cell culture, algae or bacteria) without or with soft walls Does not require freezing and thus may avoid artefacts of freezing Requires many expensive machinery

14. CRYOGENIC GRINDING (Grinding in Liquid Nitrogen) Used for hard tissues and cells like roots, stems, but also for hard walled cells like some algae and cyanobacteria Low temperature protects the proteins during grinding Time consuming and requires suitable machinery

15. LYSIS BUFFER Used only for bacteria or animal cells May cause degradation No machinery needed

16. THANKYOU