HI All,
My team is looking at the changes in distribution of the OTR in a range of male reproductive tissue. Currently getting non-reproducible results for RAT testis and hippocampus positive controls. We are using parafilm embedding and citric acid antigen retrieval methods.
If you have experience in this area some helpful suggestions would be greatly appreciated.
best
mike
Hi Michael, it depends of your used antibody. First check the data sheet and look weather the antibody is suatable for the kind of application you are using (formalin fixed, paraffin embedded (ffpe). Second weather a antigen rietrieval is genrally recommanded. If not , don't do it. Try to find out your optimal antibody concentration for your dedection and staining system e.g. labelled secondary antibody with fluorophore or peroxidase. If the concentration would be to high take a more sensitive detections system like biotinylated secondary + StreptAvidin-fluorophore or peroxidase. You can also try to make a antigen retrieval in EDTA-buffer pH 9. Check the optimal concentration in the same way I have discribed it abough. Good luck!
Immunohistochemistry (IHC) utilizing a monoclonal antibody provides specific localization of OTRs in the human brain and thereby provides opportunity to further study OTR in human development and psychiatric conditions.
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