serum cytokine detection - (Sep/14/2012 )

Hi everyone,

I'm new to serum sample ELISA and if anyone can kindly help me, I'd really appreicate it.

1. Can I use DuoSet from R&D to detect IFN-g and IL-4 in mouse serum? Or do I have to use specific kits that are designed for serum samples?
2. If specific serum kits are needed, do you have any recommendations? Anyone has experience with ebioscience products?


http://www.ebioscien...et-go-elisa.htm

http://us.ebioscienc...c=United States

3. I'm wondering whether I have to dilute my serum sample. The cytokine level might be low so that I can't dilute it. If not diluted, the serum matrix is very complex and different from that of the reagent diluent used to dilute the standards. Are these two background comparable?

Thanks in advance

I have used the eBioscience kits for a wide variety of cytokines though for cell supernatants. They work very well and I used to do multiple serial dilutions where possible- like undiluted, 1:10 and 1:100. If you know that the cytokine levels are going to be low, you can just use undiluted samples and not waste reagents.

I haven't used Duoset but I guess the brochure does say that it is for cell sups and that you have to optimise it for sera.But I guess it should work with sera too.They also have a Proteome profiler array which will help you look at a variety of cytokines and chemokines if you are interested.

http://www.rndsystem...rraypanela.aspx

I used (tried) ebioscience to measure c-Jun in cell culture. It was a nightmare and I had no success. I was rushing when I ordered the kit and didn't appreciate that it was for "qualitative analysis". Although if you're just glancing at their protocol they try and make it look like you'll be able to set up a standard curve, which is not case.

If you have the option to go with R&D, whom I have had great success in cell culture with, I would recommend their kit.

Ebioscience may have more developed kits for what you're trying to measure, just be vigilant and vet their protocol thoroughly before spending $500 on something that may be a waste of your time.