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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)

Preparation and use of stock solutions It is very convenient to have stock solutions of the main salts and buffers used during purification. Correctly and carefully prepared stock solutions can noticeably improve accuracy and reproducibility of the purification procedures. Stock solution preparation sequence

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