Immunoelectrophoresis

Immunoelectrophoresis (IEF) is a method for studying the antigenic composition of biological materials, combining electrophoresis and immunodiffusion . First described by Grabar and Williams in 1953 , in 1965 the method was modified by Scheidegger to minimize it (the so-called micromodification of the IEF method).

A sample of antigenic material is separated by gel electrophoresis (usually agarose ), resulting in the formation of characteristic zones. Then, precipitating antiserum is introduced in parallel with the electrophoresis zones, antigens and antiserum diffuse towards each other, and in the place where the antiserum meets the antigen, precipitation lines appearing in the form of an arc . After immunodiffusion and elution of the non-precipitating molecules from the gel, the gel is stained with special dyes (for example, amide black 10B , azocarmine B and other dyes that stain proteins in the case of protein antigens or Sudan black B in the case of lipoprotein antigens). There are also a number of modifications of the IEF method (using pure antigen, using a monospecific antiserum, the IEF method according to Osserman, the IEF method according to Geremans) ^[1]