Timing for soluble protein purification procedure

Protein crystallography Programs for crystallography X-ray detectors
Basic tutorials:
Protein Peptide

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

Step-by-step protein purification timing

Preparation of the crude extract	1 hour
Ion-exchange chromatography on DEAE-Sepharose Fast Flow	1-2 hours
SDS-PAGE (optional)	1.5 hours
Ammonium sulphate cut or preparation of sample for hydrophobic chromatography in 1.5M - 1.8M AmSO₄	0.5-1 hour
Hydrophobic chromatography on Butyl, Phenyl or Ether-Toyopearl 650S	1-2 hours
Concentration on VivaSpin concentrators or Ammonium sulphate precipitation for storage or to prepare a sample for gel filtration	0.5-1 hour
Gel filtration	2 hours
SDS-PAGE analysis	1.5 hours
Concentration and preparation for storage or use	1 hour

The whole procedure takes from 5 hours (for two steps procedure) to 13 hours (for a 3 step procedure with SDS-PAGE after first step)

Navigation: Go back to the Protein purification
Navigation: Go back to the Chapter 3. Common sense
Navigation: Go to: previous Purification of DNA-binding proteins
Navigation: Go to: next Preparation of the stock solutions