Faculty of Biological Sciences

Protein purification using FPLC

FPLC using an ÄKTA Explorer

This chromatography system is most used for purifying over-expressed, recombinant proteins. Performing gradient elution (rather than step elution) and combining separations based on different protein properties gives a better product.

We can do affinity chromatography ( NTA, GST etc.), ion-exchange, hydrophobic interaction and gel filtration chromatography.

Ask us about designing strategies to give purer protein than using an affinity tag alone.

Monitoring elution at up to three wavelengths (in the UV and visible spectrum) helps in selecting protein versus nucleic acid peaks or dye-labelled protein.

Fractions can be collected into different volume tubes or into microtitre plates.

NTA purification protocol (pdf)
About ÄKTA protein purification (pdf)

Protein over-expression and purification

Ask us about methods and materials for protein over-expression and purification.

We have plasmids for expression of His-, GST and MBP-tagged proteins. We also have competent cells for a range of strains, to address a range of protein over-expression problems. We can also give you a set of standard methods – good starting points if you are over-expressing and purifying a protein for the first time.

Insoluble protein purification and refolding protocol (pdf)
Large-scale protein expression protocol (pdf)