On my proposal research I wrote that in vivo anti-inflammatory effect will be determined in plasma and peripheral white blood cells by using ELISA method. I want to try other methods for cytokines estimation.
You can try at the transcription [m-RNA] level using specific probes. lalitha kabilan
You could examine acute phase protein levels in the plasma. The selection of these proteins will be species dependent. APP are more long lived than cytokines in the plasma and there are good commercial ELISA kits available.
Hi All, thank you for the answers.
The transcription level/qPCR sounds great. I have thought about doing PCR for it, but i can't do it now since there's no facilities in my place. I am trying to look for other place.
@Prof.Carolyn : Is the APP estimation proper to my research? My research focus is inflammation in gout. So, I want to know the profile of cytokines especially IL-1 since we know IL-1 is a crucial mediator of gouty inflammation.
Western blot sounds good. It is affordable for me at this time.
Western blot is not used often for cytokine detection so you need to be sure that any antibodies you use are qualified for that application. Also remember that IL-1 beta needs to be cleaved to be released and detected by Elisa so RNA analysis may not always correlate with the Elisa.
Dear Dr. Young,
Thank you for your kind help to answer my question. Can you tell me how to cleave IL-1 beta? Is it a compulsory step to detect IL-1 beta by ELISA? because i have run my in vitro assay for IL-1 beta without any other process. Thank you in advance.
IL-1 is cleavage is complex, see minireview at http://www.rndsystems.com/mini_review_detail_objectname_MR99_IL-1.aspx. It is not thought to be released without cleavage but I don't know if an uncleaved IL-1 beta is ever just shed from cells (e.g. in exosomes).
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