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Dialysis into Lipid


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#1 TomH

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Posted 26 May 2010 - 07:40 AM

Hi all,

I have a membrane protein currently solubilised in detergent and I want to replace the detergent with lipid, and have been told the best way to do this is via a slow dialysis. Unfortunately I have been unable to find any protocols on actually how to do this. Can anyone point me in the right direction?

#2 sgt4boston

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Posted 27 May 2010 - 02:41 AM

I believe lipids are insoluble in water. It may be best to first remove the detergent from your protein by dialysis in the final buffer you require using tubing with MW cut off smaller than the protein and add in the lipid post dialysis.

Hopefully, after removal of the detergent your protein will not fall out of solution. If the lipid is soluble and MW is less than that of the protein and can pass thru the tubing pores you could use the lipid buffer as your dialysis buffer.

thoughts?

#3 TomH

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Posted 27 May 2010 - 04:06 AM

My protein will def fall out of solution without detergent. I was told I could dialyse my protein / detergent solution against a lipid solution and over time the detergent molecules would be replaced by the lipid molecules. It's just the detail on how to actually do I was looking for.

I assume the lipids would be soluble, or at least form soluble entities (liposomes or something?) so they would be able to be dialysed in solution.




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