Hi Mahmoud,

Lipids can bind to proteins and reduce their protein solubility. Together with the salt content, they will affect the current in IEF.

Sample clean-up is thus necessary. You can try using TCA/acetone precipitation or 2D-clean-up kit to remove salts and lipids.

Beware not to over air-dry the protein pellet after the solvent removal. Good luck.

Lipids should be removed. Because they interfere with IEF. You can use TCA/acetone precipitation to remove lipids and salt.

Hi Mahmoud,

Lipid components interfere with 2D electrophoresis, so better option is to carryout protein precipitation to separate non protein molecules from the mixture. Usually we use acetone:ethanol:acetiacid in the ratio of 50:50:0.1 solution followed by low speed centrifugation to separate protein from the mixture.

All the best

Hi,

our protein service lab prefers the 2-D clean-up kit from GE Healthcare. We tried other kits and methods too, but this one gives the best results for 2D-gels.

All the best, Kristin