AW: [ccp4bb]: Lipid/LPS contamination

[Alexander.Pautsch@bc.boehringer-ingelheim.com]

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dear Konstantinos

you can specifically stain  LPS on gels according to Tsai&Frash 1982
Anal.Biochem. 119, 28199. And afterwards the proteins w. comassie as usual.
We got  rid of LPS through ion exchange (or chromatofocussing) on a Mono-P
or Resource -Q. If you are dealing with a membrane protein you could
optimize the detergent extraction step.
Another strategy is to express the protein into inclusion bodies (virtually
LPS free) and refold afterwards, this method of course has some limitations.

Regards
Alex 

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Dr. Alexander Pautsch
 Protein Crystallography /Structural Research 
 Boehringer Ingelheim Pharma KG Deutschland 
 Birkendorferstrasse 65
 88400 BIBERACH, Germany 
 tel. +49 - (0)7351 - 54 4683 
 fax. +49 - (0)7351 - 54 98390 
 email alexander.pautsch@bc.boehringer-ingelheim.com
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> -----Ursprüngliche Nachricht-----
> Von:	Konstantinos Beis [SMTP:kb11@st-andrews.ac.uk]
> Gesendet am:	Freitag, 14. Dezember 2001 17:53
> An:	ccp4bb@dl.ac.uk
> Cc:	awa@dl.ac.uk
> Betreff:	[ccp4bb]: Lipid/LPS contamination
> 
> ***  For details on how to be removed from this list visit the  ***
> ***          CCP4 home page http://www.ccp4.ac.uk         ***
> 
> Dear All,
> I am sorry to bother you with a non CCP4 related email. I am working with
> a
> protein and I am afraid that I have lipid/lps contamination of the 'pure'
> protein solution. I cannot use amm. sulph. cut since my protein is
> crashing
> out. I can remove some of them with DEAE-cellulose column and
> ultracentrifuge. At the moment I am assessing for LPS/lipids with silver
> stained gels. Do you know any other method(s) that can check for lps?
> Thanks
> in advance.
> 
> 
> Konstantinos Beis
> 
> 
> ____________________________________________________________________
> Konstantinos Beis                               Email:
> kb11@st-andrews.ac.uk
> Centre for Biomolecular Sciences                Tel  :+44 1334 4272 65
> University of St. Andrews
> North Haugh
> St. Andrews
> FIFE KY16 9ST, UK
> 
> WWW: http://speedy.st-andrews.ac.uk/~kostas/kostas.html
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