It can be seen from the formula of electrophoretic mobility that there are many factors affecting the separation of electrophoresis. Some of the main influencing factors are briefly discussed below: 1. The nature of the biological macromolecule to be separated The charge, molecular size and nature of the biomolecule to be separated will have a significant effect on electrophoresis. In general, the larger the charge amount of the molecular band, the smaller the diameter, and the closer the shape is to the spherical shape, the faster the electrophoretic migration speed. 2. The nature of the buffer The pH value of the buffer will affect the degree of dissociation of the biomolecule to be separated, and thus affect its charging properties. The farther the pH value of the solution is from its isoelectric point, the greater the net charge is, and the speed of electrophoresis is also The larger, especially for amphiphilic molecules such as proteins, the pH of the buffer also affects the direction of electrophoresis. When the pH of the buffer is greater than the isoelectric point of the protein molecule, the protein molecule is negatively charged and the direction of electrophoresis is directed to the positive electrode. In order to maintain the stability of the biomolecule to be separated during electrophoresis and the pH of the buffer, the buffer usually maintains a certain ionic strength, generally between 0.02 and 0.2. If the ionic strength is too low, the buffering capacity is poor, but if the ion is If the intensity is too high, a strong oppositely-charged ion diffusion layer (ie, an ionic atmosphere) is formed around the molecules to be separated. Since the ionic atmosphere and the molecules to be separated move in opposite directions, electrostatic attraction is generated between them, thereby causing electrophoresis. The speed is reduced. In addition, the viscosity of the buffer also affects the electrophoresis speed. 3. Electric field strength The electric field strength (V/cm) is the potential drop per centimeter, also called the potential gradient. The higher the electric field strength, the faster the electrophoresis speed. However, increasing the electric field strength causes an increase in the current intensity through the medium, which causes an increase in the amount of heat generated by the electrophoresis process. The work done by the current in the medium (W) is: W=I2.Rt In the above formula: I is the current intensity, R is the resistance, and t is the electrophoresis time. Most of the work done by the current is converted to heat, which causes the temperature of the medium to rise, which has many effects: 1 sample and buffer ion diffusion rate increase, causing the sample separation band to widen; 2 convection, causing mixing of the object to be separated; 3 if the sample is sensitive to heat, causing protein denaturation; 4 causing medium viscosity reduction, resistance drop, etc. . The heat generated in electrophoresis is usually emitted from the center to the outer periphery, so the temperature of the center of the medium is generally higher than that of the outer circumference, especially the tubular electrophoresis, thereby causing the viscosity of the central portion of the medium to decrease relative to the peripheral portion, the friction coefficient to decrease, and the electrophoretic migration speed. Increasing, since the electrophoresis speed of the central portion is faster than the edge, the electrophoretic separation band is usually bow-shaped. Reducing the current intensity can reduce the heat generation, but it will prolong the electrophoresis time, causing the increase of the diffusion of the biological macromolecule to be separated and affecting the separation effect. Therefore, in the electrophoresis experiment, the appropriate electric field strength should be selected, and the temperature can be appropriately cooled to obtain a better separation effect. 4. Electroosmosis The relative movement of a liquid in an electric field to a solid support medium is called electroosmosis. Since there may be some charged groups on the surface of the support medium, such as the surface of the filter paper usually has some carboxyl groups, the agar may contain some sulfate groups, and the surface of the glass usually has Si-OH groups and so on. After ionization of these groups, the surface of the supporting medium is charged, and some oppositely charged ions are adsorbed, and moved toward the electrode under the action of the electric field to form a flow of the solution on the surface of the medium. This phenomenon is electroosmosis. When the pH value is higher than 3, the surface of the glass is negatively charged, and the positive ions in the solution are adsorbed, causing the solution layer near the surface of the glass to be positively charged. Under the action of the electric field, it migrates to the negative electrode, causing the electrode solution to generate electroosmotic flow to the negative electrode. . If the direction of electroosmosis is the same as the direction of electrophoresis of the molecule to be separated, the electrophoresis speed is accelerated; if not, the electrophoresis speed is lowered. 5. Support mesh holes The mesh size of the supporting medium has a significant effect on the electrophoretic migration rate of the isolated biomacromolecules. The swimming speed is fast in the medium with large mesh holes, and the swimming speed is slow. As a high-tech enterprise, Shengchuang Biotech Co., Ltd. will make unremitting efforts to develop into a biological company specializing in the sales and service of molecular biology reagents, protein purification reagents and biomedical instruments. Shengchuang Biotechnology Co., Ltd. focuses on the frontier and development of life sciences, with "Quality First, Customer First" as the company's values, and the introduction of "new products, new technologies" as its mission, focusing on providing professionalization for Chinese researchers. Product and technical services. 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