During processes like protein extraction and purification, often the cells are subjected to break down. These broken cell structures, called lysed cells when contained in a fluid is called protein lysate.
In order to use the protein lysate later in various experiments, as and when required, it is preserved after the process of extraction. Therefore, it becomes necessary to keep the protein extracts in proper condition, without compromising on the quality and functionality of the ingredients of the cell.
This is because the degraded cells are likely to affect the results of the experiments in the laboratories. Thus, the preservation of the protein lysate becomes necessary so that they do not hamper the results due to the degraded cell components.
Ways to preserve the protein lysate
- Cold environment
After the lysis of the cells, the proteins start to degrade, and this shortens their lifespan. Therefore, you need something that will either stop or reduce the denaturation of the protein lysate.
The most effective way to keep them is in a cold place, preferably in a cold solution. This, in turn, will keep the cells in proper shape and keep the quality. The buffers, detergents or salt, whichever is used for lysis, should be cold.
- Careful handling
The whole process of extracting the cell organelles is a tedious task; therefore, handling of the cells, as well as its components, is the key. This will make sure the retention of active and functional cytoplasm in the protein lysate.
The process of extraction depends on the type of experiment you want to use it for. The extraction procedure might seem to be violent, yet you should handle the buffers carefully.
The two ways to get protein lysate are from the cell cultures or the tissues. Generally, a protease such a trypsin is used during lysis, and they tend to degrade the quality of the cell structures largely, which is utterly undesirable.
Therefore, it becomes imperative that you rinse off that extra trypsin before rupturing the cells (lysis).
- Use buffers
It is preferable to use buffers when you want to extract the cell components. The buffers help in minimizing the damage caused to the protein lysate after the extraction of the same.
Selection of the proper buffer is also important, given the type of cells you are using for your experiment. For instance, a mammal cell, or a plant cell or a yeast, etc; all need a different kind of buffer.
The proteases used for cell lysis should be fresh. Generally, the proteases are enzymes and they tend to lose their properties over time. That is why they need to be fresh to function properly.
Often these inhibitors are available in concentrated form; during this situation, you may dilute them and use for your protein extraction. This will keep the cell organelles of the protein lysate in shape and their properties intact.
Briefly, high-quality protein is essential for conducting effectual experiments. Therefore, the above-mentioned points can make sure proper maintenance of the same.