How to remove LPS from protein solution - (Dec/13/2005 )
Dear all
I havea problem in the LPS removal from proteins expressed in E. coli periplasm.
The proteins were constructed to a Strep-Tag (8 aa). After affinity chromatography, there is a lot of LPS in purified protein, which is 50,000 times more than dd water.
Polymixin B does not work. How about organic solvent, detergents?
Could you give me some advice?
Thank you very much.
-plymouth-
QUOTE (plymouth @ Dec 13 2005, 06:22 PM)
Dear all
I havea problem in the LPS removal from proteins expressed in E. coli periplasm.
The proteins were constructed to a Strep-Tag (8 aa). After affinity chromatography, there is a lot of LPS in purified protein, which is 50,000 times more than dd water.
Polymixin B does not work. How about organic solvent, detergents?
Could you give me some advice?
Thank you very much.
I havea problem in the LPS removal from proteins expressed in E. coli periplasm.
The proteins were constructed to a Strep-Tag (8 aa). After affinity chromatography, there is a lot of LPS in purified protein, which is 50,000 times more than dd water.
Polymixin B does not work. How about organic solvent, detergents?
Could you give me some advice?
Thank you very much.
Do I have just the thing for you! There is a little column called EndoTrap and you can purchase from Cambrex (they were originally from Germany!). They aren't cheap, but boy, did they do the trick for me! I can reuse them about 10 times, even though they say no more than 5.
Good Luck,
Cindy
-biokmst-
TRY to filter Through activated char coal, make sure that ur protein should be below one PI.
-kasa-