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.-Spectroscopy and tell you about the precautions for protein 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.
When pursuing any kind of protein purification, we must take time to protect its stability and protect its activity. There are some general considerations that need to be remembered. They include:
1. The operation should be placed on ice as much as possible or in a cold storage.
2. Don't be too dilute, the protein concentration should be maintained at μg / mL ～ mg / mL.
3. Appropriate pH, unless it is used for aggregation chromatography, the pH of the buffer solution used is the same as that of pI, preventing protein accumulation.
4. Use protease inhibitors to prevent the degradation of policy proteins by proteases; when purifying proteins in cells, participate in DNase to degrade DNA and prevent DNA contamination of proteins.
5. Prevent repeated freezing and thawing of the sample and vigorous agitation to prevent denaturation of the protein.
6. The composition of the buffer solution simulates the intracellular environment as much as possible.
7. Participate in 0.1 ~ 1mmol / LDTT (dithiothreitol) (or β-mercaptoethanol) in the buffer solution to prevent protein oxidation.
8. Add 1 ~ 10mmol / LEDTA metal chelating agent to prevent heavy metals from damaging the policy protein.
9. Use a sterilized solution to prevent the growth of microorganisms.
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