I would like to examine the level of TNF-alpha, IL-6, Il-1beta, Il-8 in the supernatant of human adipose tissue after various periods of storage in various preservative fluids. Is it necessary to prepare a sample before the detection? So far, I have been trying to harvest a supernatant and store it in -80 degrees after spinning it (from tissue contamination), or not. In both cases, the detection was placed at the lower threshold; everything shows that it should be much higher. I have used a Human Inflammatory Cytokine Cytometric Bead Array (CBA) - I Kit (RUO) Bioscience. What can I do to get some more distinct results?
Dear Natalia!
Please You look at folloowing link:
https://www.merckmillipore.com/RU/ru/product/MILLIPLEX-MAP-Human-Adipocyte-Magnetic-Bead-Panel-Endocrine-Multiplex-Assay,MM_NF-HADCYMAG-61K?ReferrerURL=https%3A%2F%2Fwww.google.ru%2F
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1534009/
You should determine where the problem is coming from. Are your "preservative fluids" ruining the cytokines? (feel free to give details -- they matter!) Or is there some other problem in your test procedure? You can reveal at what stage(s) it's going wrong by mixing authentic cytokines in known amounts into the samples at various stages during processing. That should give you some idea where to start troubleshooting.
Hi Natalia, we have looked into the possible production of cytokines by adipose tissue. This largely reflects a culture/incubution artefact with massive amounts of IL-6 formed. Yet, assessment of cytokine release in vivo (over the human omentum (visceral fat)) does not indicate cytokine release. Article Human visceral adipose tissue and the plasminogen activator ...
other groups came to similar conclusions for subcutaneous fat. In order to set the debate with regard to a possible role of TNFalfa in the metabolic syndrome we performed an intervention trial with anti-TNF. Again no signs for a role of TNFalfa in the human metabolic syndrome.Article Chronic TNF-α Neutralization Does Not Improve Insulin Resist...
so think very carefully about your research question.- Badges
- Science topic