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Fully automatic protein purification instrument - general procedure steps of fully automatic protein purification
2020-08-01
Proteins are complex biological macromolecules that play important roles in living organisms. They perform numerous tasks within the cell, involved in the structural development and location, function and regulation of tissues and organs of an organism. For the analysis of individual proteins and protein complexes, the purification of proteins by fully automatic protein purification instruments is the cornerstone of these studies. The general procedure for the separation and purification of a specific protein by automatic protein purification equipment can be divided into three steps: pretreatment, coarse fractionation, and fine fractionation. _Imported protein purification instrument_protein purification equipment_protein purification system_automatic protein purification equipment
Fully automatic protein purification equipment to separate and purify a certain protein, the first thing is to release the protein from the original tissue or cell in a dissolved state and maintain the original natural state without losing biological activity. For this reason, it is necessary to choose an appropriate method to break up tissues and cells according to different situations. Common methods for breaking bacterial cell walls include ultrasonic crushing, sand grinding, high-pressure extrusion, or lysozyme treatment. After the tissues and cells are disrupted, choose an appropriate buffer to extract the desired protein. Insoluble matter such as cell debris was removed by centrifugation or filtration. _Imported protein purification instrument_protein purification equipment_protein purification system_automatic protein purification equipment
If the desired protein is mainly concentrated in a certain cell component, such as nucleus, chromosome, ribosome or soluble cytoplasm, etc., they can be separated by differential centrifugation with an automatic protein purification instrument, and the cell component is collected as the next step Purified material. If the desired protein is bound to the cell membrane or membranous organelle, the membrane structure must be depolymerized by ultrasonication or detergent, and then extracted with an appropriate medium. _Imported protein purification instrument_protein purification equipment_protein purification system_automatic protein purification equipment
After the protein extract is obtained, an appropriate method is selected to separate the desired protein from other miscellaneous proteins. Generally, methods such as salting out, isoelectric point precipitation and organic solvent fractionation are used for the separation of this step. These methods are characterized by simplicity and large processing capacity, which can not only remove a large amount of impurities, but also concentrate the protein solution. Some protein extracts have a large volume and are not suitable for concentration by precipitation or salting out. Ultrafiltration, gel filtration, freeze vacuum drying or other methods can be used for concentration. _Imported protein purification instrument_protein purification equipment_protein purification system_automatic protein purification equipment
After the sample is roughly fractionated and separated by an automatic protein purification instrument, the volume is generally small, and most of the impurity proteins have been removed. For further purification, chromatography methods are generally used, including gel filtration, ion exchange chromatography, adsorption chromatography, and affinity chromatography. If necessary, electrophoresis, including zone electrophoresis and isoelectric focusing, can also be selected as the final purification step. _Imported protein purification instrument_protein purification equipment_protein purification system_automatic protein purification equipment
Crystallization is the final step in the separation and purification of fully automated protein purification instruments. Although the crystallization process does not guarantee that the protein is uniform, crystals can only form if a certain protein is predominant in the solution. The crystallization process itself is accompanied by a certain degree of purification, and recrystallization can remove a small amount of entrapped protein. Since denatured protein is never found during the crystallization process, crystallization of the protein is not only a sign of purity, but also a strong indicator that the product is in its native state. _Imported protein purification instrument_protein purification equipment_protein purification system_automatic protein purification equipment
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