Hi
I am trying to labelled murine CD8 LT by immunofluorescence on thymus and tumor sections embedded in paraffin and dewaxed. I meet some difficulties. I tried several Antobodies without success. Someone would have experience in the field and could help me by giving me the references of a working Ac and specify the protocol if it contains a particular point. Thank you very much.
Yannick
Hi Yannick, I had a good experience with Mouse-on-Mouse (M.O.M) Basic kit (from Vector Laboratories) some time ago when the only good antibody available was on raised in mouse and I worked on murine material. I have checked on their website and the sample microphotographs are IF with virtually no background. It is a ABC type kit containing a mouse IgG blocking agent.
To have IF detection you would need to use fluorochrome-labelled avidin or streptavidin as a last step.
GeneTex has a convincing microphotograph on the spec sheet for their mouse anti-CD8 [LT8] antibody with DAB as chromogen. You have a good chance to use this Ab, or another mouse Ab, the Vector kit and AV or Strep/fluorescent tag.
Good luck with your project.
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