Good Buffer Solutions can greatly reduce the pH fluctuation range when adding a small amount of acid or base and water, a mixed solution of weak acid and its salt (such as HOAc and NaOAc), a mixed solution of weak base and its salt (such as NH3·H2O And NH4Cl) etc. are all buffers. Buffers are widely used in biochemical experiments and play an important role in the process of protein separation and purification.
The separation and purification of protein is a complex task, and the purification process of each protein is not exactly the same. Therefore, only by using appropriate protein purification methods in the protein extraction and preparation process can a stable protein be obtained. Throughout the process, the stability of the protein depends on the multi-component buffer system used. The best condition is to dissolve the protein while maintaining biological activity.
Since most of the protein purification is performed in vitro, the buffer systems corresponding to recombinant proteins on the market can mimic the conditions of natural proteins in vivo in order to achieve the purpose of stable protein storage and transportation.
The main function of the buffer is to provide the most suitable environment for the target protein. In the process of selecting and preparing buffers, the following factors must be considered:
1. Buffer composition:
The buffer component of the buffer itself should not interact with the protein. For example, some enzymes are inhibited by the phosphate group of phosphate buffer, which can lead to loss of protein function. It also shows that the suitable buffer for each protein is not exactly the same, so users should try to choose the buffer recommended in the manual when dissolving the protein to ensure the stability of the protein and the corresponding activity. The typical buffer concentration ranges from 20 to 100 mM, and it may be necessary to add other components to ionic strength-sensitive proteins or metal ions such as magnesium (salt solution is 150 mM), which is necessary for some enzymes to be active .
2. pH:
The pH depends on the actual application of the protein. If protein is used for certain types of biological assays, such as enzyme assays, the pH at which the enzyme shows the best activity should be selected. When preparing proteins for purification techniques (ion exchange, gel filtration, etc.), the pH should be selected according to the experimental conditions to obtain the maximum purification efficiency. When a specific pH is not required, the pH at which the protein exhibits the best stability is the best. This is especially true for protein storage.
Desheng specializes in the research and development, production and sales of blood collection tube additives, in vitro diagnostic reagents, biological buffers, and luminescent substrates. A variety of buffer materials (TRIS, HEPES, MOPS, etc.) can be provided, and buffers of different concentrations can be configured according to customer needs.
