The method of protein purification, separation and purification of protein, the protein is released from the original cell or tissue in a dissolved state and remains in the original natural state without loss of biological activity. For this reason, the material should be free of animal and fatty tissue contamination, the seed material should be even decontaminated from the seed coat by substances such as tannins, and oil seeds preferably use low-boiling organic solvents.
1. Protein purification method, size and different separation and analysis methods: dialysis and ultrafiltration use these proteins to pass through the properties of molecules that cannot pass through semi-permeable membranes; density gradient centrifugation The quality and density of proteins have a greater influence on particles when they are centrifuged in the medium Settling faster; gel filtration a column chromatography.
2. Separation of solubility differences in use: Electroprecipitation Since the net charge of protein molecules is zero at the isoelectric point, the electrostatic repulsion between molecules is reduced, easy to aggregate, and the minimum solubility; salt solution used for salting out with salt solution The concentration increases or decreases the solubility of the protein.
3. The charge is separated by different analysis methods, the main contents include electrophoresis and ion exchange chromatography.
4. Protein purification method The selective adsorption and separation of proteins utilizes the strength of particle adsorption.
5. Affinity Chromatography Ligand-Based Separation This biological property allows protein molecules to bind specifically rather than covalently to another molecule called a ligand.
6. Protein purification method Low-temperature organic solvent precipitation method: Use water-miscible organic solvents, such as methanol and ethanol, to reduce the solubility and precipitation of most proteins. This method is higher than resolution salting out, but protein denaturation is more severe When easy, it should be done at low temperature.