Abstract
Publisher Summary This chapter describes isoelectric focusing (IEF) in liquid media, polyacrylamide gels, or sephadex, utilized both analytically and preparatively to fractionate, purify, and evaluate the structure and/or genetics of many different immunoglobulins. Several reviews have described the methodology and application of liquid and gel IEF of immunoglobulins. Analytical IEF is performed in polyacrylamide gel. This technique became a frequently used method for studying the heterogeneity of anti-body responses and screen genetically defined populations of laboratory animals for the pattern of specific antibodies. Analytical polyacrylamide gel IEF can be performed in two ways—in narrow tubes or in a thin layer on a glass plate. The specific detection of antibodies in thin-layer gel IEF extended considerably by overlaying the gel with antigen. Detection of the antibody-bound antigen or substrate is based on (1) the use of radioactive antigen and autoradiography, (2) the use of the enzyme activity of the antigen, and (3) the use of antigen bound to erythrocytes and subsequent complement-dependent lysis.
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