Currently, I am studying about the inflammasome activity inhibition using a compound in cultured microglial cell lines. As the common procedure to analyze inflammasome activity is by measuring the secreted IL-1B using ELISA, I would like to know whether we could analyze by measuring the downregulation of inflammasome related genes such as Nlrp3, Asc, Casp1, Il1b, and Il18. Is it possible?
The short answer is yes. However, a decrease in basal levels of mRNA encoding for Nlrp3, Asc, Casp1, Il1b, and Il18 could be informative when it is not accompanied by cell death following any treatment.
I would add Dr.'s Divaker Choubey's answer, the suggestion to perform Western Blotting of the inflammassome proteins to be able to compare mRNA levels with protein expression levels.
I believe it would give a robustness to the inhibitory response.
Yes, real time PCR would help in assessing the mRNA expression of the above mentioned genes; in most cases, I prefer to perform the mRNA analysis before proceeding to the next step. You could further corroborate the mRNA data by performing protein expression analyses (ELISA, Western etc) or you may look for the most relevant functional phenotype associated with inflammasome. This would be only to help you tie togetehr each of your findings in a sequential manner such that ultimately your overall results are well supported. Best regards
Thank you very much for all of your suggestion Divaker Choubey , Marcel Rodrigues Ferreira , Rashmi Pathak
- Badges
- Science method
- Similar topics
- Medicine
- Immunology
- Immunology Techniques
- Immunoassay
- ELISA