Yes you can before (baseline values) and after (acute or chronic) stress treatment. You shall always try to optimize your methodology and use highly sensitive methods (ELISA is one of the commonly immunological methods we use but you also can assessed the cytokine levels by other molecular methods such as real-time RT-PCR and western blot, dot blots, arrays...).  You can determine the levels both at RNA and protein level and study their expression regulation etc etc.

Yes you csn ,you can use ELISA method to do it. 

Although cellular mRNA expression of all cytokines can be quantified, but not the plasma protein level is naive animals. The levels of some cytokines like TNF & IL-1is very less in normal animals & they need some induction stimulus to get released into the circulation. However, the levels of some constitutively expressed cytokines like IL-6 is detectable in plasma of healthy animals.

Thanks to all of you,

I have done a little trial using MSD assay (IFN-alpha, IL1-B, IL-2/4/5/6/8/10/12, TNF-alpha). Unfortunately the Abs that I used are specific for mice, I work with rats (better than nothing). I got good levels of TNF and IL-6 from after a week from the stress.

Thanks,

Matteo

Of course you can use any GOOD ELISA kit, and as you have mentioned there is cross reactivity between mice and rats, but use kits specific to RATS only. My student used one from BD biosciences some years back that worked fine.

Hi Mattteo,

The detection and quantification of equine cytokines has been hampered by the lack of antibodies for many years. But now Luminex came up with a fluorescent bead-based system that allows the simultaneous detection of the cytokines in a single sample (you cam measure up to 200 cytokines/analyte). So you can save cost of individual cytokine ELISA kit and cost-effective tool for quantification of various cytokine. I have used it, its really good and mainly 10 microliter of sample is sufficient.

I hope it may help you. You can go to Millipore web site and you will get Rat abs ELISA kit.

All the best.

Regards

Ramanatha

Actually, I ordered the Meso Scale Discovery assay for rat TNF-alpha and IL-6. Not so expensive and really sensitive 1pg/ml. Let's see how it goes.

Thanks to everyone,

Matteo

MSD plates should work well. I am also collecting samples from mice (serum) to run the assay on plates from MSD.

Matteo, Can you update how well it works in your case? 

Good luck!

Janak

Hi,

to determine TNF alpha levels before and after in vitro stimulationwith mitogen LPS  is a useful method to ensure reliable measurement of cytokine levels.

Have a nice weekend !

Regards,

Andreas

Hi there, we have measured cytokines using a multi-plex elisa from Bio-rad. They have a rat specific one which is great and you only need 10ul plasma. We got the 24 plex (so 24 different cytokines measured in the same same). Cost is quite high £2000 but definitely worth it if you can.

Besides ELISA, you can also use quantitative mass-spectrometry for this purpose.

Arseniy Yuzhalin, would you recommend any article on citokine levels/ quantitative mass-spectometry?

Thanks