We suspect that the 'ferocious' immune response that we observe is due to the presence of a conjugated ligand.
Hi Sharon,
What exactly do you mean by conjugated? If you're referring to a specific protein interaction between, for example, a receptor and ligand, or multiple receptors, you can try a proximity ligation assay (PLA). PLA is an immunofluorescent technique where you label a section with two antibodies and attach DNA probes to them. When there is close proximity of both antibodies, the DNA probes can be ligated and the DNA can be replicated by rolling circle. After this has been done, a fluorescent probe is added to the mix, and you see fluorescent spots on your slide where there is sufficient proximity.
More info here:
https://www.ncbi.nlm.nih.gov/pubmed/23921974
http://www.bio-protocol.org/e1479
Hope this helps,
Scott
Hello,
Fixing the cells, immunofluorescent staining with specific antibody and fluorescence microscopy. The protocol optimized for your cells and protein of interest is very important, it should be searchable.
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