[Histonet] RE: Antigen retrieval
TGoins <@t> mt.gov
Wed Jun 13 10:13:19 CDT 2012
A good retrieval method will restore the epitope to a configuration that the antibody will recognize. The data sheet received with a commercial antibody may suggest a retrieval method to use, but if you are using a non-commercial source, trying a variety of methods may help unless the epitope is irreversibly altered by fixation.
We run all new assays using pH 9, pH 6 and sometimes pH 3 [10 min treatment rather than 20 min] in addition to enzymatic retrieval using Pepsin, Trypsin or Protease and finally No Retrieval.
Does anyone have a favorite antigen retrieval method for FFPE mouse tissues that they would be willing to share? I have been using citrate buffer Ph6.0 with poor to moderate results. Thanks for any help!
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