Detecting this lipid by antibody is maybe unusual, but for me a very interesting idea, since this is a strong immunogen. PAGE is not used for lipids, even if you will separate it by acrylamide, would rather not transfer to the membrane. ELISA works well for water-soluble chemicals, not the lipids. But why not to try the Thin-Layer-Chromatography (TLC) to separate it and develop with antibody as in the classical Western-Blot (WB)? You will have the hydrophobic solvent mixture for your lipid separation, in the water buffers for WB your lipid should not dissolve. Only make sure your stationary phase in TLC will not dissolve in water. If this will work for lipids, you will have a very nice Nature/Science article;)
Thank you for your insight. What do you think the monoclonal could attach to since there is no longer an O-antigen as on the intact LPS molecule?
Please don't focus on monoclonal antibodies only. Polyclonal can recognize it as well. The size of your molecule is huge, with the proteins/peptides I'm more familiar, than lipids, it is known 3 amino-acids could be recognized by the antibody. Suggest you to flag your question with lipid/immunology topics rather than sds-page or western, since there you will have better expertise.
Couldn't lipid A BE detected with the HEk blue cell based assay, or radioimmune assays found on the internet. Like LPS, LIPID a is cytotoxic.
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