BIOLOGY 2B03 Lecture Notes - Lecture 6: Gel Electrophoresis, Affinity Chromatography, Ion Exchange
27 Sep 2016
School
Department
Course
Professor
Document Summary
How to purify proteins: develop a good assay, select protein source, break open cells-protein extract, solublize protein: in vitro, aqueous, stabilize protein: keep in native form, fractionate, determine purity, protein assay a way of monitoring your protein. Method of determining the presence of a specific protein and estimating the concentration of the protein. Should be based upon a unique characteristic of the protein of interest. Ex: znymatic activity, immunological assay(antibody), biological activity/effect: protein source should be. Easy obtainable in large amounts (e. g. tissue, cultured cells, production in e. coli, yeast) Contain a high concentration of the protein of interest. Low in proteins that may co-purify, containing similar properties. Low in proteases that may destroy the protein of interest: lyse open the cells, protein solubilization. Soluble proteins some cytosolic proteins, serum proteins secreted proteins: hydrophilic r groups. Insoluble proteins transmembrane or membrane associated proteins aggregate in aqueous solutions: hydrophobic r groups. Protein solubility is affected by ph, detergents, salt concentration: protein stabilization.
