BIOLOGY 2B03 Lecture Notes - Lecture 5: Transmission Electron Microscopy, Scanning Electron Microscope, Biological Membrane
Document Summary
Why purifying proteins: 2 types of scieniic analysis - strutural, funcional, backwards to determine aa sequence gene seqeunce, develop anibodies. Ways you can do this - change ph, slat concentraion, detergent: stabilize protein (maintain naive structure) Maintain noncovlalent interacions by - temperature (hot=degradaion), protease inhibitors, ligands, Concentraion of salts, metal ions, concentraion of protein, ph: all maintain conformaion, fracionate (separate you protein from all other ones) size, polarity, charge, solubility, shape have to do muliple fracionaions, determine purity. Diferenial centrifugaion - separate based on size mass and density. Many interacions with beads - protein is going to move slowly vice versa - able to difereniate base on what comes out of column. Ion exchange chromotography - posiively charged beads/negaively charged proteins if beads and protein are opposites they sick - use nacl to disrupt charge and make protein low can also use warm water soluion or change ph of a soluion.
