High definition chromogenic multiplex IHC staining using UltraPlex technology allows precise visualization of tissue biomarkers. Our rapid staining technology and simple workflow using cocktails of antibodies and a single antigen retrieval step allows you to go from staining to analysis in a single day. Our range of BOND RX/RXm autostainer-optimized staining panels provides a seamless, high-throughput solution for biomarker tissue profiling.
Routine multiplex immunohistochemistry detection has been limited to two chromogens using primary antibodies from different species detected using enzyme-labeled secondary anti-species antibodies followed by chromogen precipitation. Cell IDx’s UltraPlex hapten technology overcomes many of these limitations allowing detection of 3–4 markers using primary antibodies from any species while employing a single antigen retrieval step on an autostainer.
To demonstrate that the UltraPlex IHC and UltraPlex IF technologies are detecting the same biomarkers, two serial tonsil slides were incubated with the same hapten-labeled primary cocktail and one slide was developed chromogenically and the other fluorescently using their respectively-labeled anti-hapten antibodies. The results are presented below using the PD-L1/CD8/GranzymeB panel.
The UltraPlex multiplex IHC protocol requires 4.5 hours for 3-marker staining, as illustrated below. The initial step in this protocol is identical to Cell IDx’s multiplex immunofluorescence protocol in which a cocktail of hapten-labeled primary antibodies are incubated on the tissue. For chromogen detection, enzyme-labeled anti-hapten antibodies are employed instead of fluor-labeled anti-hapten antibodies. See protocol schema in Technology page.