Immunoelectrophoresis
Immunoelectrophoresis is a laboratory method used in biochemistry, immunology and other fields of science. This method combines two techniques, electrophoresis and immunodiffusion, to identify and measure proteins in a sample.
Overview
Immunoelectrophoresis is used to separate proteins based on their electrophoretic mobility and their reaction with specific antibodies. The process begins with electrophoresis, which separates proteins in a sample based on their size, shape, and electrical charge. After electrophoresis, the proteins are made to react with specific antibodies in a process known as immunodiffusion. This results in the formation of visible precipitin lines, which can be analyzed to identify and measure the proteins.
Applications
Immunoelectrophoresis has a wide range of applications in various fields of science. In clinical laboratories, it is used to diagnose and monitor diseases that affect protein levels in the body, such as multiple myeloma and autoimmune diseases. In research laboratories, it is used to study the properties of proteins and their interactions with antibodies.
Procedure
The procedure for immunoelectrophoresis involves several steps. First, a sample containing proteins is applied to a gel and subjected to electrophoresis. This separates the proteins based on their electrophoretic mobility. Next, the gel is overlaid with a layer of agarose containing specific antibodies. The proteins diffuse into the agarose and react with the antibodies, forming visible precipitin lines. These lines are then analyzed to identify and measure the proteins.
Limitations
While immunoelectrophoresis is a powerful tool for protein analysis, it has some limitations. The method requires specific antibodies for each protein of interest, which can be expensive and time-consuming to produce. Additionally, the method is not suitable for analyzing complex mixtures of proteins, as the precipitin lines can overlap and become difficult to interpret.
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Contributors: Prab R. Tumpati, MD