Jul 04, 2024
Version 2
TDP-43 RNA aptamer staining to detect pathological TDP-43 in FFPE human tissue, as described in Spence and Waldron et al., 2024 (Acta Neuropathologica): A SOP and tick-sheet. v2. V.2
- 1University of Aberdeen
Protocol Citation: Fergal M Waldron, Holly Spence, Jenna Gregory 2024. TDP-43 RNA aptamer staining to detect pathological TDP-43 in FFPE human tissue, as described in Spence and Waldron et al., 2024 (Acta Neuropathologica): A SOP and tick-sheet. v2.. protocols.io https://dx.doi.org/10.17504/protocols.io.eq2lyjo4mlx9/v2Version created by Jenna Gregory
Manuscript citation:
References for citation of this method
Please cite both of these below if using this method:
The citation, Spence and Waldron et al., 2024, for the first publication for the development, modification and employment of theTDP-43 RNA aptamer to stain human tissue published in Acta Neuropathologica.
RNA aptamer reveals nuclear TDP-43 pathology is an early aggregation event that coincides with STMN-2 cryptic splicing and precedes clinical manifestation in ALS.
Holly Spence*, Fergal M. Waldron*, Rebecca S. Saleeb, Anna-Leigh Brown, Olivia M. Rifai, Martina Gilodi, Fiona Read, Kristine, Roberts, Gillian Milne, Debbie Wilkinson, Judi O’Shaughnessy, Annalisa Pastore, Pietro Fratta, Neil Shneider, Gian Gaetano Tartaglia, Elsa Zacco, Mathew H. Horrocks, Jenna M. Gregory‡. Acta Neuropathologica 2024 Mar 5;147(1):50. DOI: 10.1007/s00401-024-02705-1. *equal contributions, ‡corresponding author.
The citation, Waldron and Spence et al., 2024, for this SOP published on protocols.io is:
TDP-43 RNA aptamer staining to detect pathological TDP-43 in FFPE human tissue, as described in Spence and Waldron et al., 2024 (Acta Neuropathologica): A SOP and tick-sheet. v2.
Fergal M. Waldron*, Holly Spence*, Jenna M. Gregory‡. protocols.io 2024;
DOI: dx.doi.org/10.17504/protocols.io.eq2lyjo4mlx9/v2. *equal contributions, ‡corresponding author.
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: March 01, 2024
Last Modified: July 04, 2024
Protocol Integer ID: 102860
Keywords: ALS, MND, FTD, ALSFTSD, TDP-43, RNA aptamer, Human tissue staining, Immunohistochemistry
Funders Acknowledgement:
Target ALS
Grant ID: BB-2022-C4-L2
Disclaimer
Authors declare no conflicts of interest.
Abstract
Here we provide a SOP to outline the correct procedures for performing Immunohistochemistry (IHC) to detect pathological TDP-43 in FFPE-preserved human tissue using the TDP-43 RNA Aptamer, as described in Spence and Waldron et al., 2024 published in Acta Neuropathologica (DOI: 10.1007/s00401-024-02705-1).
Users with access to Sequenza immunostaining racks and histological facilities (with fume hood) should be able to carry out all steps over two days.
This protocol uses the TDP-43APT sequence published in Zacco et al., 2022. The sequence is: CGGUGUUGCU with a 3' Biotin-TEG modification, purified using HPLC, scale: 1.0 µM synthesis.
References for citation of this method
Please cite both of these below if using this method:
The citation, Spence and Waldron et al., 2024, for the first publication for the development, modification and employment of theTDP-43 RNA aptamer to stain human tissue published in Acta Neuropathologica.
RNA aptamer reveals nuclear TDP-43 pathology is an early aggregation event that coincides with STMN-2 cryptic splicing and precedes clinical manifestation in ALS.
Holly Spence*, Fergal M. Waldron*, Rebecca S. Saleeb, Anna-Leigh Brown, Olivia M. Rifai, Martina Gilodi, Fiona Read, Kristine, Roberts, Gillian Milne, Debbie Wilkinson, Judi O’Shaughnessy, Annalisa Pastore, Pietro Fratta, Neil Shneider, Gian Gaetano Tartaglia, Elsa Zacco, Mathew H. Horrocks, Jenna M. Gregory‡. Acta Neuropathologica 2024 Mar 5;147(1):50. DOI: 10.1007/s00401-024-02705-1. *equal contributions, ‡corresponding author.
The citation, Waldron and Spence et al., 2024, for this SOP published on protocols.io is:
TDP-43 RNA aptamer staining to detect pathological TDP-43 in FFPE human tissue, as described in Spence and Waldron et al., 2024 (Acta Neuropathologica): A SOP and tick-sheet. v2.
Fergal M. Waldron*, Holly Spence*, Jenna M. Gregory‡. protocols.io 2024;
DOI: dx.doi.org/10.17504/protocols.io.eq2lyjo4mlx9/v2. *equal contributions, ‡corresponding author.
Image Attribution
Jenna M Gregory
Guidelines
This protocol is designed for users with access to Sequenza immunostaining racks and histological facilities (with fume hood) should be able to carry out all steps over two days.
Before start
Please see appendix for materials required to carry out this protocol.
Materials
See SOP appendix for materials.
Safety warnings
Safety First
Before starting, please ensure all relevant Health & Safety documentation is in order including the following
- COSHH assessment
- Risk assessment
- Safe System of Work
- SOP read and understood
Before start
Please see appendix for materials required to carry out this protocol.
Protocol references
References for citation of this method
Please cite both of these below if using this method:
The citation, Spence and Waldron et al., 2024, for the first publication for the development, modification and employment of theTDP-43 RNA aptamer to stain human tissue published in Acta Neuropathologica.
RNA aptamer reveals nuclear TDP-43 pathology is an early aggregation event that coincides with STMN-2 cryptic splicing and precedes clinical manifestation in ALS.
Holly Spence*, Fergal M. Waldron*, Rebecca S. Saleeb, Anna-Leigh Brown, Olivia M. Rifai, Martina Gilodi, Fiona Read, Kristine, Roberts, Gillian Milne, Debbie Wilkinson, Judi O’Shaughnessy, Annalisa Pastore, Pietro Fratta, Neil Shneider, Gian Gaetano Tartaglia, Elsa Zacco, Mathew H. Horrocks, Jenna M. Gregory‡. Acta Neuropathologica 2024 Mar 5;147(1):50. DOI: 10.1007/s00401-024-02705-1. *equal contributions, ‡corresponding author.
The citation, Waldron and Spence et al., 2024, for this SOP published on protocols.io is:
TDP-43 RNA aptamer staining to detect pathological TDP-43 in FFPE human tissue, as described in Spence and Waldron et al., 2024 (Acta Neuropathologica): A SOP and tick-sheet. v2.
Fergal M. Waldron*, Holly Spence*, Jenna M. Gregory‡. protocols.io 2024;
DOI: dx.doi.org/10.17504/protocols.io.eq2lyjo4mlx9/v2. *equal contributions, ‡corresponding author.