I am trying to measure expression of TLR3, TLR7 and TLR9 by flow cytometry. I know they are all located in the endosomal wall looking inside. Does anybody have experience if regular intracellular staining protocols after cell permeabilization allow to measure proteins in this localization as well?
This is a detailed intracellular staining protocol. Hope to help you.
https://www.creative-bioarray.com/support/detection-of-cytokines-by-flow-cytometry.htm
most intracytoplasmic labeling methods are applicable to visualize TLRs (from 2 to 10), whether present in early or late endosomes. The simplest and least expensive technique is the use of the final 0.3% saponin to permeabilize the cells without prior fixation. Note: If you are studying pathological cells, it is important to look for TLRs also on the surface of the cell. You could be very surprised!
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