Imprint immunofixation of electrofocused immunoglobulins

An imprint immunofixation (IIF) technique for the characterization of electrofocused immunoglobulins (Ig) is described. Electrofocused proteins are blotted (imprinted) from the separating polyacrylamide gel to agarose gels by gel-to-gel overlays. The protein imprints are chemically fixed in the agarose gels with a solution of 46% methanol, 8% acetic acid and 46% water. The imprinted Ig are then identified radioimmunologically, using an indirect system with monoclonal mouse anti-human Ig antibodies in the first layer and 125I-labelled rabbit anti-mouse Ig in the second, followed by autoradiography. The method is sensitive and permits characterization of Ig in unconcentrated cerebrospinal fluid. By sequential imprinting, each separated specimen can be characterized for up to 10 separate antigenic determinants without loss of sensitivity.