LPT 321: Meat Science (1+1)

ELECTROPHORETIC METHODS

• The various electrophoretic methods include
• Agar gel electrophoresis
• Starch gel electrophoresis
• Polyacrylamide gel electrophoresis
• Sodium dodecyl sulfate polyacrylamide gel electrophoresis
• Iso-electric focussing
• Agar gel electrophoresis
• Electrophoretic separation of proteins on the basis of mobility of proteins in a supporting gel of agarose at a constant pH and electrical field is termed as Agar gel electrophoresis.
• Counter Immuno Electrophoresis (CIE)
• The principle of this technique is very similar to agar gel immunodiffusion.
• The diffusion of antigen and antibody is facilitated by the application of a low voltage current. Further, the endosmosis created by the agar gel changes the charge of g -globulins and moves them towards cathode.
• The meat proteins moving towards anode provide immunoprecipitate with homologous g -globulin at the point of equivalence.
• Polyacrylamide gel electrophoresis (PAGE)
• Polyacrylamide gels are prepared using acrylamide and bis which provides cross linking between the polymerized long chains of acrylamide.
• The separation of proteins on the basis of their mobility in a polyacrylamide gel is comparatively better since the resolution of different proteins are optimum. Polyacrylamide gel electrophoresis also requires a constant pH and electrical field to provide high resolution of protein.
• Iso-electric focussing (IEF)
• It is an electrophoretic technique which utilizes the charge at the surface of the protein to drive it through a gradient gel.
• The pH gradient is setup by polyacrylamide compounds, the ampholytes.
• The proteins applied on to the gel reach a point where the surface charges becomes neutral at their iso electric point