I plan to measure TNF-a, IL-1B, IL-6, IFN-gamma and MCP-3. Which method should I use? Which methods are convenience, reliable, easy and cost effective?
It depends on your concentrations, how many cytokines your testing, your purity, your equipment and your price range. However, in general you're describing an ELISA plate, they're convenient, easy, and usually $150-$500 depending on the complexity of your plate. I would recommend you look into some of the ELISA plate arrays. They're premade and usually fairly inexpensive, relatively.
A multi-analysis using ELISA plates is convinient and eventually less expensive than if you used individual ELISA plates specific for each inflammatory factor. Some companies are able to customize ELISA plates.
Cytokine numbers, your purity and concentrations and etc play an important role in your measurement. I am unanimous with suggested comments. Some of the ELISA plate arrays are convenient provided that you consider whatever suggested.
http://www.rndsystems.com/product_detail_objectname_duoset.aspx
These are pretty cheap and easy to use...
If you would like to meassure small amounts of a panel of different cytokines in the same probe, a cytokine bead array (works with a flow cytometer) is a good way…
All the suggestions are good but I do want to add a caution in that you should make sure that the cytokines you are testing are standardized based on WHO international cytokine standards. If not, you will not be able to compare your results to those that other publications have reported.
do u mean I have to use standard from WHO of each cytokine to measure the cytokine level?
If you are not sure which cytokines are produced, i think the easiest and cheapest way is to start with qRT-PCR . Once you have verified the genes are expressed, then you can go back and quantify using the techniques suggested above.
Commercial ELISA kits are simple and cheap methods. But , for analysis of cytokines, one of important issue is analyses timing, because some cytokines increase at acute period of pathologies while other cytokines increase late period of pathologies. Therefore, cytokines analyses should be performed at appopriate time for each cytokines. Otherwise you can throw money to garbage. Most ideal interval can be find from related articles with pubmed search.
My comment about WHO standards is meant to emphasize that any commercial reagent kits you utilized use WHO standards (where available) in their standardization protocols. Not all cytokines have standards but many do.
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