Chapter 2 - General sequence of protein purification

- Protein crystallography

Main steps:

   - Protein purification
     - Introduction
     - Step-by-Step
       - Equipment & Reagents
       - Stock solutions
       - Chromatographic columns
       - Crude extract
       - pre Chromatographic step
       - Chromatographic step
     - Common sense
     - Protocols
     - Charts & Tables
     - Appendix
   - Crystallisation

Special:

- Programs for crystallography
- X-ray detectors

Basic tutorials:

   - Chemistry
   - Protein
   - Peptide
   - Amino Acids

Xtal community:

- CCP4BB

Protocols and tips in protein purification or How to purify protein in one day.

First step of any experiments is the understanding of the general sequence of all major steps. You should understand in great details what you should do and in which sequence. This is very important to prepare all necessary stock solutions, to book appropriate time to work with expensive equipments, such as centrifuges, columns etc. This plan of actions will help you to save a lot of time and be prepared to some non-standard situations.

2. General sequence of protein purification procedures

Preparation of equipment and reagents
Stock solutions preparation
Preparation of the chromatographic columns
Preparation of crude extract - cell free extract or soluble proteins fraction
Pre-chromatographic steps
Chromatographic steps