Single and Double Antibody Tissue Staining (Chromogenic IHC)

Chromogenic IHC is an immunohistochemical technique that uses antibodies and a chromogen/substrate combination to visualise the presence of antigens in tissue.

Single antibody staining allows the visualisation of one antigen, whilst double antibody staining allows two different antigens to be viewed within the same tissue section.

We carry out single and double staining using the Leica BOND-RX^m and the Leica BOND-MAX Autostainers and predominantly use the two following techniques:

1. Bond™ Polymer Refine Detection Kit: This uses the chromogen DAB with the HRP enzyme which is visualised via a brown precipitate.
2. Bond™ Polymer Refine Red Detection Kit: This uses the chromogen Fast Red with the alkaline phosphatase (AP) enzyme which is visualised via a red precipitate.

For a list of antibodies we have previously optimised protocols for, see here.