Electrophoresis (Part 02) | Types of Electrophoresis | Paper Electrophoresis and its Application

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Electrophoresis (Part 1)| What is Electrophoresis |Electrophoresis Principle| Phytochemistry-Part 39 • Electrophoresis (Part ...
Electrophoresis is defined as the migration (movement) of charged particles through a solution under the influence of an electrical field , leading to their separation. It is separation technique: Electrophoresis is a technique that enables the separation and analysis of charged molecules in an electric field. It is comparatively cheaper to other separation techniques. Electrophoresis of cations (positive charge) is known as cataphoresis, while electrophoresis of anions (negatively charged particles) is known as anaphoresis.
It is one of the widely used techniques in molecular biochemistry, microbiology and biochemical research for the separation and analysis of proteins in a mixture, DNA and RNA. There are two main types or electrophoretic methods, depending upon whether the separation is carried out in the absence or presence of a supporting or stabilizing medium.
1. Free solution electrophoresis/ Moving boundary electrophoresis
a. Capillary electrophoresis
b. Iso-tachophoresis
c. Immunoelectrophoresis
d. Isoelectric focusing
2. Electrochromatography or Zone electrophoresis:
a. Paper electrophoresis
b. Gel electrophoresis
c. Thin layer electrophoresis
d. Cellulose acetate electrophoresis
Paper electrophoresis:
In this electrophoretic techniques special filter papers (Whatman filter paper No.1 or No.3) is used as supporting medium. The charge carried by the molecule depends on the pH of the medium. The mobility of the ion depends on the voltage applied and the charge on the ion.
In this technique, filter paper (supporting medium) is moistened with the buffer, immerse both the ends of filter paper in a buffer solution reservoirs containing the electrodes . The sample is applied as spots in the centre of the paper, high voltage is applied, and the spots migrate according to their charges. After electrophoresis, the separated components can be detected by a variety of staining techniques, depending upon their chemical identity .
1. This technique is useful for separation of small charged molecules such as amino acids and small proteins.
2. It is a simple and economical method.
3. Paper electrophoresis is adapted well to routine laboratory use.
4. Alkaloid identification.
5. Environmental studies
6. Serum analysis for diagnostic purpose
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