Will an antibody recommended for IHC-P with one retrieval buffer work with another?

We have performed antigen retrieval comparisons with a number of products.  We consistently see the strongest signals with high pH retrieval buffers.  We have not seen the staining pattern influenced by the choice of retrieval protocol; however, the signal intensity can be affected.  Antibodies that yield an acceptable signal with low pH retrieval (citrate buffer) will generally work well with high pH retrieval (EDTA or Tris-EDTA), but the optimal concentration may not be the same.  Antibodies that require the more robust high pH retrieval are unlikely to work as well with low pH retrieval. It is generally our recommendation to use EDTA retrieval when assembling antibodies in a multiplex panel.
 

Last updated: September 12, 2024