LPS isolation by hot phenol\water method - (Feb/11/2006 )

Hello

I really would thank somebody who send me this LPS meth, I really need it.

Please, If somebody can help me.

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hi chailie

I need the article " hot phenol water method for LPS". can you send it for me?

THANK YOU

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Dear All,

Please find attached a modified LPS extraction protocol (if you do happen to use it I would be very appreciative if you could quote the paper at the top of the protocol).

Best wishes,

Charlie.

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Thanks.

Do you know the difference between rough LPS vs smooth LPS in terms of chemical structure?

If someone is just interested in extracting lipophilic LPS, would you recommend going through sonication/ enzyme treatment etc?

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To answer your questions, some Gram negative bacteria express rough LPS (including N. gonorrhoeae) that does not express an O-antigen (it does however have a core region including KDO etc) it is also termed LOS (lipooligosaccaharide). Other Gram negative bacteria including E. coli and Salmonella express LPS that normally has an O-antigen (repeating unit of sugars) and is termed smooth (due to the appearance of the colony as grown on agar).


For extracting lypophilic LPS you can use hot phenol-water but the LPS may be found in the phenol fraction which is more complicated to purify (the protocol I sent by attachment outlines the procedure). In this case you can also try the PCP method (phenol chloroform and petrol ether). This is the favoured method used by Uli Zähringer (Borstel Centre) but like the hot phenol-water method I would recommend sonicating the bacterial solution before enzyme treatments.

Hope this helps,

Cheers,

Charlie.

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Hi All,
Can someone help me and send me the "LPS isolation by hot phenol"? please...

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Hi,everybody.
I am interested in that if the LPS Extracted by this P-W method is biologically active to raise antibody by immuning it to animals.
Anyone has proposals?

yours,
tozz

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Dear sir ,Charles D. Ellis,
Recently we have been involved in characterization phosphomannomutase a gene which affect LPS structure in many bacteria.
And hence we are looking for efficient protocol for LPS extract.
When I search on google, I found your information at http://www.protocol-online.org/biology-for...13636more1.html.
In fact I have your paper published in Mol Microbiology.It would be great help if you could able to send me detailed modified protocol for LPS extraction and also
Original paper on "hot phenol water method for LPS"Thanks in advance
Yours sincerely
veeranagouda




Dr. Y. Veeranagouda
Postdoctoral research fellow
Department of Microbiology
Changwon National University
Changwon-si, Kyoungnam 641-773
South Korea
Phone : 82-551-213-2758
Mobile : 008210 5458 1979
e-mail: veeresha_y@rediffmail.com
veereshagy@gmail.com

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Hallo Veeranagouda,

I doubt that he would read this, he has not been active since december this year and this is a very old post.
Maybe you could email him at: c.ellis@imperial.ac.uk

good luck with the research.

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hi,
Thank you so much.
I will try.

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