PXD041199-2

PXD041199 is an original dataset announced via ProteomeXchange.

Title	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
Description	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.
HostingRepository	PRIDE
AnnounceDate	2024-10-22
AnnouncementXML	Submission_2024-10-22_06:45:11.588.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Darragh O'Brien
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2023-03-30 01:22:56	ID requested
1	2024-06-15 23:05:06	announced
⏵ 2	2024-10-22 06:45:12	announced	2024-10-22: Updated project metadata.

10.1007/s00401-024-02729-7;

Ellis MJ, Lekka C, Holden KL, Tulmin H, Seedat F, O'Brien DP, Dhayal S, Zeissler ML, Knudsen JG, Kessler BM, Morgan NG, Todd JA, Richardson SJ, Stefana MI, Identification of high-performing antibodies for the reliable detection of Tau proteoforms by Western blotting and immunohistochemistry. Acta Neuropathol, 147(1):87(2024) [pubmed]

submitter keyword: antibody validation,Tau, MAPT

Darragh O'Brien
contact affiliation	Nuffield Department of Medicine. University of Oxford
contact email	darragh.obrien@ndm.ox.ac.uk
lab head
Darragh O'Brien
contact affiliation	Target Discovery Institute, University of Oxford
contact email	darragh.obrien@ndm.ox.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD041199
     1. Label: PRIDE project
     2. Name: 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