RNA/Protein Co-Detection

ACD always recommends starting by establishing a working IHC protocol and then using the same established secondary for ICW. You may use any secondary you have been using in the lab for your IHC workflow.  Please ensure that any secondary that you choose uses the appropriate detection chemistry (e.g. HRP-conjugated secondary for ICW, fluorescent-conjugated or HRP-conjugated secondary for fluorescent detection).

With the increased compatibility of the new integrated co-detection workflow across a wide variety of antibodies, there are no specific antibody clone recommendations. ACD encourages you to use your preferred clones for RNA-protein co-detection. For best results, titrate your preferred primary antibody concentrate in ACD’s Co-Detection Antibody Diluent in the context of the integrated co-detection workflow.

Co-Detection Blocker prevents cross-detection of RNAscope signal by the IHC detection steps.

The ACD Antibody diluent has been formulated to ensure maximal retention of RNA sample quality.

Our sequential dual ISH/IHC workflow allows for IHC staining following RNAscope. Protease pretreatment is necessary for RNAscope but can impact your target protein of interest. Some epitopes may be better conserved than others after protease treatment and some optimization of protease may be necessary to find an optimal condition that allows for both RNAscope and IHC detection. The new integrated co-detection workflow cross-links the primary antibody prior to the protease step, allowing conservation of the antigen/antibody for detection.

RNA/protein co-detection is compatible with the RNAscope 2.5 HD RED, RNAscope Multiplex Fluorescent V2, BaseScope v2, and miRNAscope HD (RED) assays.