Electrophoresis

by Sam Malone


Mobilization of electrically charged substance in the presence of an electric field is electrophoresis. This is attributed to the coulomb force which is in turn in correlation with the electrical conditions. Molecular biology makes use of  gel electrophoresis. This is a procedure commonly used for nucleic acid (DNA and RNA) isolation. It involves the loading of DNA, RNA and proteins on a gel, which reduces molecular mobility. This technique isolates them, based on their electrophoretic mobility. The most commonly used is the frail, large pore sized colloid, agarose gel, whose molecular mass is around 12,000.

Agarose is a linear polysaccharide isolated from seaweed and is damaged on handling. It is used primarily to detach large molecules of a molecular mass larger than 200 kdal. Though agarose gels derive faster results, their promise is substandard because of the uncontrollable pore size. It comprises of agarobiose in a repeated manner, which has galactose and 3,6-anhydrogalactose in an alternate fashion. It is usually utilized at around 1% to 3%.

Based on the frictional forces and charges capillary electrophoresis (CE) is used to isolate ionic species. This method is involves the usage of fused-silica capillaries to isolate a complex range of molecules, both large and small. The capillary tube radiates heat willingly and thus over heating of samples is prevented. Molecules are separated based on their charge, water repelling qualities and size. An assortment of separation  techniques are followed including Non-Aqueous Capillary Electrophoresis (NACE), Capillary Gel Electrophoresis (CGE) , Isotachophoresis (ITP), Electrokinetic Chromatography (EKC) Capillary Isoelectric Focusing (CIEF), Capillary Zone Electrophoresis (CZE), Micro Emulsion Electrokinetic Chromatography (MEEKC), Capillary Electrochromatography (CEC), and so on.

Diseases in humansis diagnosed by identifying specific proteins in the blood and this method is referred to as Serum Protein Electrophoresis (SPE). This procedure diagnoses amyloidosis and myeloma. Amino acids form proteins, which act as a carrier for the positive and negative electrical charge and they mobilize in fluid in the presence of an electrical field. An electrical field is required to isolate the serum proteins, based on their shape, charge and size. It is a globulin specific test. It is a laboratory procedure wherein blood is collected in a syringe. A special paper with agarose gel is taken and the serum is placed over it and kept in an electrical field. It diagnoses hypogammaglobulinemia, a disorder attributed to decreased gamma globulin levels. Such individuals are more prone to infection.

In this procedure, based on the electrical charge, serum proteins are further classified into globulin, alpha 2 globulin, albumin alpha1, beta globulin and gamma globulin.


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