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Protocols and tips in protein purification

Protocols and tips in protein purification or How to purify protein in one day Second edition 2018 Department of Molecular Biology & Biotechnology 2 3 Contents I. Introduction 7 II. General sequence of protein purification procedures Preparation of equipment and reagents Preparation and use of stock solutions Chromatography system Preparation of chromatographic columns Preparation of crude extract (cell free extract or soluble proteins fraction) Pre chromatographic steps Chromatographic steps Sequence of operations during IEC and HIC Ion exchange chromatography (IEC) Hydrophobic interaction chromatography (HIC) Gel filtration (SEC)

purification and sometimes classical methods are considered to be defunct. This is not the case. The most justified use of tags is for difficult expressions, such as expression of eukaryotic proteins in E.coli, expression of the domains of the proteins, toxic proteins, in the cases when tags help polypeptide chains to fold, in

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