Can you write a report on Protein Experiment No. (7) Electrophoresis, Title: Protein Electrophoresis Q&A please Aim of Experiment: Determination of individual protein in serum Theory: One of the simple techniques for quantitation of serum protein is their separation into an electric field and this procedure is referred to as "serum protein electrophoresis (SPE). Electrophoresis is widely used for separating protein fractions. In addition to separating albumin from globulins this technique further fractionates the globulins into four or five major group: alphal (Q), alpha2 (az) or betal, beta2 (B1, B2) and gamma (v) globulins. Electrophoresis is similar in many ways to chromatography, except that separation is due to the movement of charged particles through an electric current. If these particles are charged differently they will move in opposite directions, the positively charged particles to the cathode and the negatively charged to the anode. In this separation technique a buffered medium at a fixed pH is essential to ensure that the charge on the particles and therefore the rate of migration is stabilized. Electrophoresis is especially used for the separation of macromolecular substances such as proteins and lipids, although it is applicable to many other substances as well. In routine clinical testing the various protein substances in human serum are separated and quantitated, enzymes are separated and quantitated, enzymes are separated and identified, and various hemoglobin fractions are identified. In addition isolation and quantitation of lipoproteins can also be carried out. The above substances contain numerous fractions of similar mobility, therefore, when submitted to electrophoresis, the sharpness of separation will depend upon the extent to which each fraction is homogeneous in its migration. Each fraction carries a net charge which varies with the pH of the environment. The direction and speed of electrophoresis depends upon several factors such as (1) net charge of the molecule, (2) size and shape of the molecule, (3) electric field strength, (4) nature of the supporting medium, and (5) the temperature of operation. One of the important factors above is the pH of the buffer solution. The most commonly used pH for protein electrophoresis is pH 8.6, under these conditions most proteins are negative and therefore migrate toward the anode