I would like to perform some in vitro stimulation assays and can't find a source for L.pn. LPS. Thanks for your help.
I found the TRI reagent method in this paper to be convenient for isolating LPS from Pseudomonas aeruginosa.
Yi EC, Hackett M. Rapid isolation method for lipopolysaccharide and lipid A from gram-negative bacteria. Analyst. 2000;125:651–656
Hi Adam, thanks for this advice. Is it possible that you send me the paper you mentioned? Unfortunately, we do not have access to this Journal.
Do you know if there is the possibility to buy Legionella LPS from a company? That would be the easiest way...
Hi Geraldine, I can't send you the paper since it is not open access and not my own. I suggest you contact the corresponding author directly.
Although I could not find a vendor for Legionella LPS, I recommend you contact Prof. Dr. Helmut Brade of Glycobiotech GmbH, a company that specializes in bacterial lipopolysaccharides. It is not in the catalog, but maybe he can help you locate someone who has it, or can make it for you.
If you prefer a personal service please call us but announce your call by email:
+49-(0)-4552-1798
Prof. Dr. Helmut Brade
Fax
+49-(0)-3212-1811194
Dear You can download above mentioned paper from this website Its freely available there
http://lib3.dss.go.th/fulltext/Journal/analyst/analyst2000/no.4/4,vol.125%202000p651-656.pdf
Hi here,
For Legionella LPS or other LPS customized production you could also contact LPS-BioSciences. This is a company specialized in lipopolysaccharide production.
This is the e-mail address: [email protected]
Hope it can help somebody
Best Regards,
Tatiana
@ Geraldine I had a look at the LPS structure of Legionella pneumophila, and either phenol water or phenol chloroform extractions derived from the original Westphal method should work quite well and gives ultra high purity. For even higher purity I would dialysis the prep at the last step.
You need to be quite careful with the preparations if you are making more than small amount, the endotoxin effect is probably stronger than you would have had anticipated if you never played with purified LPS before. Rubbing your eyes with contaminated gloves or poke yourself with contaminated tips is a big NO NO.
Biotech companies generally use phenol water extractions, and the purity is achievable easily with standard protocols. The extraction only takes a couple of hours, you get very pure LPS on the next day.
I can sent you detail protocol if you are extracting.
There are other tricks to increase purity or extract deep rough LPS, but I doubt you will need those.
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