Immunoelectrophoresis. BCH To learn the technique of immunoelectrophoresis. -Technique based on the principles of electrophoresis of antigens and. Immunoelectrophoresis is a general name for a number of biochemical methods for separation and characterization of proteins based on electrophoresis and. Immunoelectrophoresis is a variation of the Ouchterlony double diffusion in gel . a two-step procedure that combines the principles of zone electrophoresis and.

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Second they require rather large amounts of polyclonal antibodies. The band begins to disappear at the 1: Polyclonal immunoglobulins are illustrated by the speckled pattern and specific protein by the solid bands. The principles underlying each method and the strengths and weaknesses of each is described.

By using this site, you agree to the Terms of Use and Privacy Policy. Each patient arc is compared with the control arc on the other side of the respective trough see Figure a for an actual illustration.


Laurell CB Quantitative estimation of proteins by electrophoresis in agarose gel containing antibodies. The immunoelectrophoretic analysis ad modum Immunoelectrophoresia is the classical method of immunoelectrophoresis.

Rocket immunoelectrophoresis is one-dimensional quantitative immunoelectrophoresis. As the antigen becomes more dilute a band is seen at the 1: This indicates the patient’s serum contains an elevated monoclonal IgG.

Other symbols and abbreviations are the same as in Figure.

The circles at immunoelevtrophoresis bottom of the gels represent control wells where antigen and antibody are placed to ensure there is a reaction. Immunoelectrophoresis differs from blotting techniques because with IE the entire procedure is conducted in an agarose gel and blotting is not necessary.

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Electrophoresis is conducted in the first stage and immunoprecipitation using antibodies against specific proteins in the second stage without removing the proteins from the separation media usually agarose. Both can be seen on the agarose gel electrophoresis SPE to the left. Also, see Figure and Figure b for a reproduction of actual gel plates. Advances in Anatomic Pathology To the left serum agarose protein electrophoresis SPE immunoelectrophoresis the migration of serum proteins.


All variants of immunoelectrophoresis require immunoglobulinsalso known as antibodiesreacting with prrinciple proteins to be separated or characterized. Immunofixation electrophoresis is largely used in medical or clinical laboratories for identifying abnormal monoclonal immunoglobulins associated with diseases of lymphocytes such as multiple myeloma. Hickman PE and Koerbin, G. The track on the left shows agarose gel serum electrophoresis SPE.

The method has been used for quantitation of human serum proteins before automated methods became available.

Key concepts Immunoelectrophoresis differs from blotting techniques because with IE the entire procedure is conducted in an agarose immunoekectrophoresis and blotting is not necessary. The introduction of the immunoelectrophoretic analysis gave a great boost to protein chemistry, some of the very first results were the resolution of proteins in biological fluids and biological extracts.

Some variants of affinity immunoelectrophoresis are similar to affinity chromatography by use of immobilized ligands.

Diagrammatic illustration of IEP. References Bence Jones H Papers on chemical pathology: The open structure of the immunoprecipitate in the agarose gel will immynoelectrophoresis additional binding of radioactively labeled antibodies to reveal specific proteins.

Chromatin immunoprecipitation Immunodiffusion Ouchterlony double immunodiffusion Radial immunodiffusion Immunoelectrophoresis Counterimmunoelectrophoresis.


Towbin H, Staehalin T and Gordon J Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets, procedures and some applications.

See Figure a and b and the text associated with it for more details of the IFE method. Today gel electrophoresis followed by electroblotting is the preferred method for protein characterization because its ease of operation, its high sensitivity, and its low requirement for specific antibodies.

The distinct band green arrow reflects a monoclonal IgG protein from a patient with multiple myeloma. Notice that the normal IgA migrates the closest to the positive electrode and the Immunoelectroporesis closest to the negative electrode. The bands represent various serum proteins, as indicated. New England Journal of Medicine The procedure used in most medical laboratories is IFE where the proteins are immunoprecipitated on the gel in a pattern that is similar to their location on routine agarose gel electrophoresis.


Affinity immunoelectrophoresis has been used for estimation of binding constantsas for instance with lectins or for characterization of proteins with specific features like glycan content or ligand binding.

Biochemistry methods Electrophoresis Molecular biology Protein methods Laboratory techniques Immunologic tests. Submit a note to the editor about this article by filling in the form below.

An electrophoresis equipment with a horizontal cooling plate was normally recommended for the electrophoresis. This page was last edited on 21 Novemberat The thick band just below the origin represents a highly concentrated monoclonal free light chain.

The abnormal protein bands identified on IFE line up with bands seen on routine agarose electrophoresis making interpretation simpler.

Migration is towards the immunoelectrophorewis electrode at the bottom. Princlple variation has been used for identification of allergens through reaction with IgE. With zero dilution, the reaction is so weak that nothing is seen. Wasting time scanning endless search results for the right article?

The sensitivity and resolving power of crossed immunoelectrophoresis immunoelectgophoresis than that of the classical immunoelectrophoretic analysis and there are multiple variations of the technique useful for various purposes. This type of immunoelectrophoreisis is easier to interpret and more sensitive than the IEP that it replaced.

Views Read Edit View history. TV, represents a trough containing trivalent antisera: On the far left, is shown agarose gel electrophoresis Age of the urine.