Dataset Information

Comprehensive Mapping of the Tau Antibody Landscape as a Resource for Western Blotting and ImmunohistochemistryComprehensive Mapping of the Tau Antibody Landscape as a Resource for Western Blotting and Immunohistochemistry

ABSTRACT: Background: The microtubule-associated protein Tau has attracted a diverse and ever-increasing research interest, with the protein being mentioned in the title/abstract of nearly 34,000 PubMed-indexed publications to-date. To facilitate studies into Tau biology and its role in neurodegeneration, a multitude of Tau-targeting antibodies have been developed, with hundreds of distinct antibody clones currently available for purchase. Nevertheless, concerns over antibody specificity and limited understanding of the performance of many of these reagents has hindered research into the protein. Methods: We have characterised the performance and specificity of 53 commercially-available Tau antibodies by Western blot, 35 of which were additionally profiled by immunohistochemistry. Antibodies were tested for their reactivity to both murine and human samples, with supporting data generated by mass spectrometry-based detection. Results: We show that presumed Tau knockout human cells continue to express residual protein, providing evidence of Tau isoforms generated by exon skipping. Our data also reveals that the binding of several antibodies presumed to detect Tau independently of any post translational modifications (i.e. “total” Tau antibodies, as well as an antibody targeting Asp421-cleaved Tau), were partially inhibited by phosphorylation. Moreover, we found that several total and phospho-Tau antibodies cross-reacted with MAP2 isoforms, while the “oligomer-specific” T22 antibody detected monomeric Tau on Western blot. With one exception, the phosphorylation-dependent Tau antibodies tested were found to not cross-react with the unphosphorylated protein. Importantly, several total and isoform-specific Tau antibodies failed to detect Tau expressed at low endogenous levels, highlighting the importance of antibody choice for studying physiological Tau expression, especially in non-neuronal cells and peripheral tissues. Conclusions: We identify Tau antibodies across all categories (total, PTM-dependent and isoform-specific) that can be employed in Western blotting and/or immunohistochemistry applications to reliably detect even low levels of Tau expression with high specificity. This work represents an extensive resource that serves as a point of reference for future studies with respect to anti-Tau antibody use and development.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

DISEASE(S): Alzheimer's Disease

SUBMITTER: Darragh O'Brien

LAB HEAD: Darragh O'Brien

PROVIDER: PXD041199 | Pride | 2024-06-16

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	20230320_HAP1_MAPT_Proteomics.xlsx	Xlsx
	QE1123_MSQ1631_20190401_DOB_TAU_A1.raw	Raw
	QE1123_MSQ1631_20190401_DOB_TAU_A2.raw	Raw
	QE1123_MSQ1631_20190401_DOB_TAU_A3.raw	Raw
	QE1123_MSQ1631_20190401_DOB_TAU_A4.raw	Raw

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Publications

Identification of high-performing antibodies for the reliable detection of Tau proteoforms by Western blotting and immunohistochemistry.

Ellis Michael J MJ Lekka Christiana C Holden Katie L KL Tulmin Hanna H Seedat Faheem F O'Brien Darragh P DP Dhayal Shalinee S Zeissler Marie-Louise ML Knudsen Jakob G JG Kessler Benedikt M BM Morgan Noel G NG Todd John A JA Richardson Sarah J SJ Stefana M Irina MI

Acta neuropathologica 20240518 1

Antibodies are essential research tools whose performance directly impacts research conclusions and reproducibility. Owing to its central role in Alzheimer's disease and other dementias, hundreds of distinct antibody clones have been developed against the microtubule-associated protein Tau and its multiple proteoforms. Despite this breadth of offer, limited understanding of their performance and poor antibody selectivity have hindered research progress. Here, we validate a large panel of Tau ant ...[more]

PMID: 38761203

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