How to remove lipids from proteins.

question: How do
remove lipids
protein when the total protein is removed from the protein? If the destination protein may be a lipid-binding membrane protein.
: Everyone who has studied lip valves knows that it is very difficult to isolate lipids from cell proteins. To use a super
centrifuge
, 100,000 or more centrifuges a few or even ten hours, a bit like the extraction of cell membrane protein. There is also a method of hierarchical extraction, which is also cumbersome. So far there seems to be no easy way to remove lipids.
: membrane protein generally has hydrophobic groups, through its water solubility to remove lipid substances. In order to improve the water solubility of hydrophobic proteins in water, it is usually able to add some urea, non-ionized me, detergents or sex ion detergents. In addition to increasing the water solubility of the protein, these substances can also remove lipids from the protein or sub-base. The solubility effect of urea is that it destroys hydrophobic and ion bonds and makes the protein partially fold. The concentration of urea depends on the requirements for dissolution and the effect of protein denaturation, generally with 4-8mol/L. urea-containing samples, in the gel should also be added to the corresponding concentration of urea.
sometimes urea can improve results by using tritonx-100 or other non-ion defiant detergents. Non-ion decontamination agent can improve the solubility of protein without destroying the structure of
protein
and maintaining biological activity, but it is necessary to choose the appropriate concentration, too low concentration can not improve the solubility of protein, too high will make the protein belt wider, affect resolution and cause texture phenomena. Glycine is a sex ion that can increase protein solubility through its even-polar moment effect. Usually with a concentration of 1%. In addition, some protein
reagents
sodium deoxycholicate added to the box also seems to remove lipids. Because we all know that bile acid can increase the water solubility of lipids, thereby removing lipid interference during electrophoresis. After the protein is extracted, the lysate is centrifuged at high speed, which also helps to remove some membrane material. It's also good to centrifuge before you go up.
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