LIFESCI 3 Lecture Notes - Lecture 8: Elution, Acrylamide, Intron
Document Summary
Small proteins enter all pores & will migrate slower through column. Larger proteins can access few or no pores & elute faster bc it can just bypass column and wash down. Separates proteins by charge (sum of charges on aa side chains) Beads are modified with + or charged chemical groups. Salt buffer is used to elute proteins that bind to charged beads. Low salt concentration: proteins that weakly bind are eluted. High salt concentration: proteins that strongly bind are eluted. Separates proteins based on affinity to bind to ligand. Immunoaffinity chromatography: uses a specific antibody for target protein, attached to column heads (highly specific) However, many proteins do not bind a ligand that can be conveniently immobilized on a column matrix. Y-shaped proteins produced & used by immune system to identify & neutralize foreign objects. Routinely used in experimental techniques of molecular biology.
