Buffer Tris in protein purification

Details:

The structure and function of protein need to be maintained in a suitable pH environment. Therefore, in the process of protein preparation and purification, the selection of biological buffer is very important. Only a suitable buffer can ensure the activity and structural stability of protein.

Buffer effect in protein purification process

There are many kinds of buffers for protein purification. While maintaining the pH of the system, it is necessary to ensure that the protein cannot be inactivated in the purification process, or to minimize the amount of protein inactivation, and to maintain the solubility and activity of the protein in each step of protein purification.

Buffer selection in protein purification process

Protein purification buffer requirements

The buffer in the process of protein purification should not only prevent protein degradation, but also prevent protein polymerization, which has a significant impact on the experimental results. The following factors should be considered in the selection of reagent and buffer: pH value, buffer system, salt ion, reducing agent and stabilizer. Each factor should be optimized according to the purified target protein, and the activity of the target protein should be used as the evaluation standard.

Understand protein stability conditions

There are many kinds of proteins, and different proteins adapt to different pH values. The buffer should be selected according to its optimal pH value. Most of the time, the pH is 7.4, which is a simulated biological condition. Most proteins are stable at this pH. However, there are some exceptions, which requires adjusting the pH value to keep the protein soluble and not degraded.

Buffer commonly used in protein purification

The commonly used buffers in the process of protein purification include Tris, Tricine, BIS Tris, mops, etc. Tris is widely used. Its buffer range is close to the physiological pH value and has low ionic strength. It is a good solvent for protein and can ensure the stability of protein. It should be noted that when preparing the buffer, try to avoid the isoelectric point pl of the target protein. Because the protein has no static charge on the surface in the pH solution of its isoelectric point, the intermolecular repulsion is reduced, it is easy to aggregate and the solubility is reduced.

Now the adaptive pH value, isoelectric point and buffer range of the protein to be purified can be found, which can be matched directly according to the corresponding requirements. Desheng is a manufacturer of in vitro diagnostic reagents, which can provide large packages of biological buffers Tris, mops, bicine, etc.