My students are going to determine the concentrations of various cytokines (e.g. TNF-alpha, IL-1beta) in the frontal cortex of mice, following drug treatment.
It is known that antidepressants such as SSRIs increase the expression of TNF-alpha and IL-1beta in the mouse brain. We do not know the exact concentration of these cytokines in the brain.
Hello,
We used Multiplex Assays for analysis of tissue homogenates which were sensitive down to a 5 micrograms/mL of protein. We were able to get moderate to high levels of inflammatory cytokines of the spinal cord or brainstem by dounce homogenizing it in 1mL of homogenate buffer supplemented with EDTA-free protease inhibitor tablets from roche. The frontal cortex would probably be equivalent in size to our brainstem sectins (maybe a little smaller). The one caveat is that we were looking at onset or peak timepoints in an adoptive transfer EAE model so there were substantial amounts of inflammation in these compartments. If you don't see substantial amounts of inflammation then I might suggest doing it in 100uL to start.
Hope this helps.
- Badges
- Science method
- Similar topics
- Medicine
- Immunology
- Immunology Techniques
- Immunoassay
- ELISA