b'Electrophoresis overview Electrophoresis is defined as the transport of chargedSupport matrixmolecules through a solvent by an electric field. Electrophoresis is a simple, rapid, and sensitiveTwo types of support matrices are commonly used in analytical tool for separating proteins and nucleicelectrophoresispolyacrylamide and agarose. The support acids. Any charged ion or molecule will migrate whenmatrices act as porous media and behave like a molecular placed in an electric field. Most biological moleculessieve. Separation of molecules is dependent upon the gel carry a net charge at any pH other than at theirpore size of the support matrix used. Agarose has a large isoelectric point and will migrate at a rate proportionalpore size and is ideal for separating macromolecules such to their charge density. as nucleic acids and protein complexes. Polyacrylamide has a smaller pore size and is ideal for separating most The mobility of a biological molecule through anproteins, peptides, and smaller nucleic acids.electric field will depend on the following factors:Polyacrylamide gel electrophoresis (PAGE) Field strength Net charge on the molecule Polyacrylamide gels are generated by the polymerization of acrylamide monomers. These monomers are crosslinkedSize and shape of the molecule into long chains by the addition of bifunctional compoundsIonic strength such as N,N-methylenebisacrylamide (bis), which react with the free functional groups of the chain termini. TheProperties of the matrix through which theconcentration of acrylamide and bisacrylamide determines molecules migrate (e.g., viscosity, pore size) the pore size of the gel. The higher the acrylamide concentration, the smaller the pore size, resulting in resolution of lower molecular weight molecules and vice versa.PAGE allows one to separate proteins for different applications based on: The acrylamide matrix Buffer systems Electrophoresis conditionsDid you knowArne Tiselius won the Nobel Prize in Chemistry for electrophoretic analysis of serum proteins in 1948.4'