For my honors thesis, I am studying the effects of social stress on Interleukin-6 levels in rats. I am planning on incubating the blood with LPS ex-vivo to simulate an immune challege and stimulate an immune response. I have ordered the LPS (E. coli prepared by phenol extraction.) I am confused as to how to add this to the blood--- the papers I have read have all said 50ng/ml of LPS, but mine came in a powder form, so I am assuming it needs to be diluted somehow? It can't mean 50ng of the power per ml of blood... I am a psychology major, and though I am very familiar with running ELISA's, I don't have anyone to ask for advice on this matter. Any help would be greatly appreciated.
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#2
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Veteran
Posted 15 October 2009 - 11:39 PM
"the papers I have read have all said 50ng/ml of LPS"
This mains you have to solve 50 ng of LPS in 1 ml milli-Q but check the leaflet that came with the LPS to confirm that this correct otherwise contact tour supplier for confirmation of the solvent.
This mains you have to solve 50 ng of LPS in 1 ml milli-Q but check the leaflet that came with the LPS to confirm that this correct otherwise contact tour supplier for confirmation of the solvent.
Ockham's razor
Pluralitas non est ponenda sine necessitate
-- "You must assume no plural without necessity".
Pluralitas non est ponenda sine necessitate
-- "You must assume no plural without necessity".
#3
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CellSpecific.com
Posted 17 October 2009 - 01:12 PM
This is a good time to read up and determine what cell subset(s) in blood will respond (produce IL-6) to LPS. Based on your findings, you might consider working with a purified cell system. Surely, through your literature reading you should be able to determine an ideal LPS concentration to stimulate IL-6 production by one or two of those subsets.
