Protein Purification System About Protein Separation and Purification Steps
When materials are pretreated and cells are crushed to separate and purify a certain protein, the protein must first be released from the tissue or cell and maintain its natural state without inactivation. So you have to break down tissue and cells in the right way. Common cell division methods include: Using mechanical shear force. This method of mechanical disruption causes the cells to rupture. General equipment, high-speed tissue grinder, homogenizer, mortar, etc.
Protein Purification System About protein separation and purification, osmotic fragmentation method This teaching method is to make cells swell and fragment under hypotonic conditions. After freeze-thawing and freezing of biological tissue, intracellular liquid ice amplifies cell bursting. Although this method is simple, be aware that those in proteins that are sensitive to temperature changes should not use this method. The ultrasonic method uses an ultrasonic oscillator to cause the uneven tension on the cell membrane to break the tissue cells. Enzymatic methods such as lysozyme destroy microbial cells, etc.
Protein Purification System About protein separation and purification, protein extraction, choose an appropriate buffer as a solvent to study the protein through extraction. The selection of environmental conditions such as the buffer used for extraction, ion strength, and composition should be based on the nature of the protein to be prepared. The membrane structure is destroyed, which is beneficial to improve the separation of protein and membrane. During the extraction process, attention should be paid to the working temperature, avoiding violent impact on stirring, etc., to prevent denaturation of the protein.
Select an appropriate method to separate the desired protein from other miscellaneous proteins. A relatively simple and convenient effective teaching method is the separation based on the difference in solubility of different proteins.
Protein purification system Regarding protein separation and purification, isoelectric point precipitation, salting out proteins to obtain proteins containing other pollutants, further separation and purification to obtain a certain purity of the sample. Commonly used purification methods include: gel filtration chromatography, ion exchange chromatography on cellulose, affinity chromatography, and so on. Sometimes these methods need to be used in combination in order to obtain a higher purity protein sample.
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