By and large ELISA is considered inferior to Mass Spec. Granted, not every one has access to this but you are going to have to validate it that way sooner or later. You may be able to use that internally but if it is critical for publication, I recommend GC/MS

If you are looking for developping sheeper EIA than individual ready to use plates, the best is to buy bulk systems. If your antibodies anti-PG anr polyclonal, get the mouse anti-rabbit for binding and the prostaglandin labeled to Acetylcholinesterase as 500 points for example and use it as bulk . Try then less amount of this tracer and optimize yuou system as well as homemade substrates and wash buffer. Of course nothing replaces GC/MS but you need to have the machine....

Michael, in agreement with previous writers, I would also recommend mass spec if you can establish a collaboration with someone with the equipment and experience, as it is clear that all RIAs/ELISAs cross react with endogenous endocannabinoid derivatives (amongst other things) and are not as specific as one would like (see PMID: 15863842). However, if this is not an option for you then I would use the Cayman Chemical EIA kit and see if you can negotiate a good bulk price. I spent years trying to couple PGE2 to various enzymes and found that it is very hard to develop an EIA for PGs that is robust and accurate; problems with non-specific binding and ligand instability made the whole exercise a nightmare. If you can't afford commercial kits than go to RIA; a much better and cheaper option for an in-house assay in my experience. I can send you a protocol if you are interested.

Thanks all for the suggestions so far, the main problem we have with MS is the cost. Until now we've been using RIAs but the cost of tracer seems to have gone up recently making them less cost effective. The info regarding cross reaction with prostamides is interesting.

Thanks again all

does anyone have a GC-MS method which is simple for prostaglandin quantitation?

If one can modify the carboxylic acid without affecting function, then you can put anything on the end such as biotin, and use streptavidin HRP, or just attach the HRP directly through standard coupling techniques that are discussed on the Internet if you look up Pierce or R and D Systems info on how they make activated HRP.

I've just received an ad from: 'Detroit R&D, Inc' about their Arachidonic acid pathway ELISA kits  www.Detroitrandd.com   [email protected] 

I'm a clinician, zero lab experience, so the only thing I can do about this is 'elucubrate'. Thanks. Salut †

I am still optimising a Gc-Ms method for detection of PGE2, n hplc, as our university has these instruments only. can someone help with a gc-ms protocol, i tried the pentafluorobromide, methoxylamine, and bstfa but only the derivitisation of mox and bsta worked up.. as for the hplc, i tried dibromoacetaphone.. it goesnt quite derivitise it. if someone an help please. The elisa kits are not as sensitive as these methods, but the elisa kits are really expensive for PGs , supplier cayman chemicals

I received an ad from: 'Detroit R&D, Inc', offering ELISA kits for Arachidonc Acid pathway. I've never participated in research lab activities, so I can't say more.

 could you please provide more information on morrow, the full name. the ultimate article goes back to pubmed which i dont have full access to the full article. and they re not on research gate either