Isoelectric focusing has been used to demonstrate the microheterogeneity of immunoglobulins. The technique is very useful when combined with immunoblotting this enables the precise antigen-binding profile of an antiserum or monoclonal antibody to be established. Immunochemists often use IEF for the analysis of a variety of antigens and preparations. This is frequently due to variation in the glycosylation of a protein and, in particular, the number of sialic acid residues present. a single primary structure which is modified after synthesis. The basis of the slight differences in the pI of isoforms as revealed by IEF is often post-translational modification of a protein, i.e. Focusing is used for the diagnosis and study of some genetically transmitted disorders. Enzymologists have used IEF to detect isoforms of purified enzymes and protein biochemists have demonstrated ‘multiple forms’ of many proteins which appear homogeneous using other biochemical techniques (including other electrophoretic methods). However, the great resolving power of IEF makes it ideal for detection of microheterogeneity in purified proteins. Brasher, Robin Thorpe, in Encyclopedia of Immunology (Second Edition), 1998 Uses of IEFĪnalytical IEF can be used to assess the complexity of protein extracts and to identify components of interest.
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