CD3, CD5 Parrafin section immunohistology.
Dear Colleagues, I have also problem in Immunostaining of Parrafin sections of Rat sciatic nerve and spleen with CD3 and CD5. I am also using rabbit anti-human CD3e, #A0452, Dako as primary antibody and using Heat method for antigen retrival, but still i am unable to establish T cell staining. Please can any body gives some suggestion to improvise my stainings. Thanks
I think you have to find anti-CD3 and anti-CD5 antibodies reactive against CD3 and CD5 of Rat. If you do not find specific antibodies for rat proteins, I suggest to try polyclonal anti- CD3 and CD5 antibodies raised against mouse or human proteins. Good Luck
well, i have tried polyclonal anti- CD3 raised human. (rabbit anti human for CD3) and for CD5 i used mouse anti rat primary antibody but i didn't get any signal.
Your antigen retrieval method can be determinant for the quality of your immunostaining.
As an example, I recommend this paper:
https://www.ncbi.nlm.nih.gov/pubmed/22821668
(Enhanced Antigen Retrieval of Amyloid β Immunohistochemistry)
Of course, as mentioned above, your antibody could have some difficulties to recognize a rat protein. Did you try to make a dot plot to test your antibody against your protein? (Could be challenging if your proteins aren't widely expressed)
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