Does anyone have a successful protocol for staining cell surface proteins of whole mount pancreatic islets? I'm mostly interested in two aspects of the process:
1. If, which, and how much detergents should be used during the procedure to limit epitope loss. 2. How should the islets be fixated (which fixative, how long) to avoid epitope masking.
Thanks a lot in advance.
Unless there is a compelling reason to do so, I would avoid using detergents when looking to stain surface proteins. This will ensure that any antibody binding you do see will be on the surface, rather than in the cytoplasm etc. Using a detergent will confound any findings even if all surface proteins of interest are not sheared. Stick with PBS+BSA/serum for blocking/staining. Good luck!
Elliot
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