[Histonet] Need for antigen retrieval question...

[Rene J Buesa]

Glenn:
  The tissues fixed in formalin undergo protein crosslinkage and need antigen retrieval (HIER).
  In a regular IHC procedure you apply the primary antibody trying to reveal the antigen epitope that has been crosslinked by using HIER. In your experiment the epitope should heve been detected  in vivo but now you are trying to detect the antibody bonded to the antigen epitope but both of them (antigen and antibody) are proteins that have been crosslinked by the formalin, and you have to "unravel" them during HIER.
  René J.

Glenn Krasinski <glenn_krasinski <@t> yahoo.com> wrote:
  My boss will be injecting mice burdened with solid tumors with antibodies specific to potential biomarkers. The tumors/tissues from the sacrificed animals will be fixed in formalin and embedded in paraffin blocks. Sections will then be stained with secondary antibodies to the specific biomarker antibodies. My question is whether or not I will need to go through an antigen retrieval process.

My apologies for the complicated question.

Glenn M. Krasinski
San Diego, CA 92121



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