assess the levels of multiple cytokines - (Dec/30/2003 )

I'd like to assess the levels of multiple cytokines from my samples, obtained from macrophages in culture.
One possibility was to use a method requiring "luminex" system. Unfortunately, I don't have access to such a device.
I wouldn't like to acquire a full ELISA kit for each cytokine...
Do you know something like a usual ELISA kit but that could be customized?

thank you for the answers

HI there!

We routinely do what you're up to, i.e. determinig the levels of various cytokines in cell culture supernatants...
In principle, there are some different approaches to your problem. One is using different kits for cytokine ELISA. ELISA kits (just the antibody pairs) is the cheapest way to measure cytokines, but i understand you do may not have that many samples to measure, and (our) kits are usually designed for about 20 96 well microtiterplates. ...
The luminex system has some advantages, but is relatively expensive. But maybe you can contact the company that offer the luminex system you've in mind and ask if they do assasy on demand, i.e. you send them the samples and the money and they send you back the data. And you do not need to buy the machine. I sure know it worked for us once....
Maybe you should consider another possibilty to determine cytokine levels, which would be by quantitative PCR, such as taqman or lightcycler. I know some companies are offering kits for murine and human cytokine panels....

Mike

If you have access to a flow cytometer then you could use the Becton Dickinson cytokine bead assay (CBA). We have had good results from just 50-100ul serum (measuring 6 cytokines at once). http://www.bdbiosciences.com/pharmingen/pr...uct.php?keyID=9
It is expensive (about £600) but it works out cheaper than doing 6 ELISAs and the software will do all of the calculations for you.

Lottie

Sorry, but the BD CBA is MUCH more expensive than even six ELISAS...

We use antibody-pairs, which are about 800 Euro for 20 96 well plates....
Makes (roughly) 40 Cent per sample per cytokine or 40 Euro per 96 well plate. So 96 sapmles with six cytokines each come for ~240 Euro... compared to 600 Pounds (~ 900 Euro).... allowing 60 Euro for additional material such as plates buffers, substrate etc. the CBA is three times more expensive!

Not that I'm saying it's a bad thing, don't misunderstand me. It's a fine system, but it's expensive, in my oppinion

Mike

I do agree mike, you can get more tests for your money from an ELISA, the CBA is a nice system if you have the money (we have negotiated discount with BD).
We have found that using 50ul aliquots of serum or supernatant instead of the 100ul recommended in the instructions gives the same result so twice as many samples can be done. it is still expensive though, although i find that it takes less time to do than running 6 ELISAs it is ideal if you have very small quantities of valuable sample.