Here we provide a SOP to outline the correct procedures to dual stain FFPE human tissue for, 1) pathological TDP-43 protein, along with 2) any other protein of interest, with the aim of visualising protein of interest histology in the context of TDP-43 pathology.
This protocol can be implemented for dual staining of any rabbit or mouse primary detection antibody (using the Novolink Polymer Detection System), with our TDP-43 RNA aptamer (biotinylated) as published in Acta
Neuropathologica here https://link.springer.com/article/10.1007/s00401-024-02705-1 in Spence and Waldron et al., 2024. The resulting dual stain will show brown chromogen (detecting the target of the primary antibody) with red chromogen (detecting TDP-43 pathology with our TDP-43 RNA aptamer.
Reference for citations of this method
Clinicopathological analysis of NEK1 variants in amyotrophic lateral sclerosis.
Olivia M. Rifai, Fergal M. Waldron, Danah Sleibi, Judi O’Shaughnessy, Danielle J. Leighton*, Jenna M. Gregory*‡ (2024). Brain Pathology doi: 10.1111/bpa.13287 (in press at the time of publication of this SOP) *equal contributions, ‡corresponding author.
Antibody and TDP-43 RNA aptamer dual staining to detect patterns of co-pathology in FFPE-preserved human tissue, as described in Rifai et al., 2024 (Brain Pathology): An SOP and tick-sheet.
Fergal M. Waldron, Olivia M. Rifai, Jenna M. Gregory‡, (2024). Protocols.io 2024; doi: tbc