Protein separation purification is widely used in the research and application of biochemistry and is an important operational skill. SCG Protein Purification System Co., Ltd.-Spectra Instruments and everyone talk about the method of protein purification and separation. A typical eukaryotic cell can contain thousands of different proteins, some are very rich, and some contain only a few copies. In order to study a certain protein, it is necessary to first purify the protein from other protein and non-protein molecules.
1. Electrophoresis method: under the same pH condition, various proteins are separated due to different mobility in the electric field due to different molecular weight and charge quantity. It is worth noting that isoelectric collection electrophoresis uses an amphoteric electrolyte as a carrier. During electrophoresis, the amphoteric electrolyte constitutes a pH gradient that is gradually added from the positive electrode to the negative electrode. The pH of the electrical point is interrupted, and this method can be used to analyze and prepare various proteins.
2. Ion communication chromatography: ion communication agents include cationic communication agents (such as: carboxymethyl cellulose; CM-cellulose) and anion communication agents (diethylaminoethyl cellulose), when the protein solution flows separately When passing through an ion communication chromatography column, the protein with the opposite charge to the ion communication agent is adsorbed on the ion communication agent, and then the adsorbed protein is eluted by changing the pH or ionic strength.
3. Affinity chromatography
Affinity chromatography is an extremely useful method to separate proteins. It often requires only one step of treatment to separate a certain protein to be purified from a very messy protein mixture with high purity.
This method is based on the fact that certain proteins can bind specifically to another molecule called a ligand (Ligand) rather than covalently.
Its basic principle: Proteins exist in a messy mixture method in tissues or cells. Each type of cell contains thousands of different proteins, so the proteins are an important part of biochemistry. Or a set of ready-made methods can remove any kind of protein from the messy mixed protein, so often adopt several methods in combination.
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