Protein purification procedures. Preparation of equipment and reagents

- 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.
Preparation of equipment and reagents
For protein purification you need the equipment and reagents listed below:
Sonicator or French Press
Centrifuge, medium speed (30-70Kg) (J-20, Avanti J-25), Appropriate centrifugation tubes
Chromatographic system comprising of, as a minimum, pump and fraction collector (normally system also includes UV monitor and recorder)
Gradient mixer
Spare pump or second chromatographic system
Chromatographic columns packed with different types of matrixes
Spectrophotometer UV/VIS (190-800nm) or VIS (340-800nm)
Bio-Rad Protein Assay Reagent and Plastic cuvettes 1.6 ml
Stock solutions of buffers components
Concentrators (VivaSpin 20 and 6, microcone) and low speed refrigerated centrifuge
Filter Device such as Filter Holder or Stericup Filter Unit.
Apparatus and solutions for SDS-PAGE
Refractometer (a pocket one for sucrose is fine)
Pippetors, tips, eppendorf tubes, tubes for fraction collector