A validated antibody panel for the characterization of tau post-translational modifications
Abstract Background Tau is a microtubule-binding protein, which is subject to various post-translational modifications (PTMs) including phosphorylation, methylation, acetylation, glycosylation, nitration, sumoylation and truncation. Aberrant PTMs such as hyperphosphorylation result in tau aggregatio...
Main Authors: | , , , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
BMC
2017-11-01
|
Series: | Molecular Neurodegeneration |
Subjects: | |
Online Access: | http://link.springer.com/article/10.1186/s13024-017-0229-1 |
id |
doaj-437f516a84194ce1aac4371077d7c061 |
---|---|
record_format |
Article |
spelling |
doaj-437f516a84194ce1aac4371077d7c0612020-11-25T00:53:00ZengBMCMolecular Neurodegeneration1750-13262017-11-0112111910.1186/s13024-017-0229-1A validated antibody panel for the characterization of tau post-translational modificationsEbru Ercan0Sameh Eid1Christian Weber2Alexandra Kowalski3Maria Bichmann4Annika Behrendt5Frank Matthes6Sybille Krauss7Peter Reinhardt8Simone Fulle9Dagmar E. Ehrnhoefer10BioMed X Innovation CenterBioMed X Innovation CenterBioMed X Innovation CenterAbbVie GmbH&Co KGBioMed X Innovation CenterBioMed X Innovation CenterGerman Center for Neurodegenerative Diseases (DZNE)German Center for Neurodegenerative Diseases (DZNE)AbbVie GmbH&Co KGBioMed X Innovation CenterBioMed X Innovation CenterAbstract Background Tau is a microtubule-binding protein, which is subject to various post-translational modifications (PTMs) including phosphorylation, methylation, acetylation, glycosylation, nitration, sumoylation and truncation. Aberrant PTMs such as hyperphosphorylation result in tau aggregation and the formation of neurofibrillary tangles, which are a hallmark of Alzheimer’s disease (AD). In order to study the importance of PTMs on tau function, antibodies raised against specific modification sites are widely used. However, quality control of these antibodies is lacking and their specificity for particular modifications is often unclear. Methods In this study, we first designed an online tool called ‘TauPTM’, which enables the visualization of PTMs and their interactions on human tau. Using TauPTM, we next searched for commercially available antibodies against tau PTMs and characterized their specificity by peptide array, immunoblotting, electrochemiluminescence ELISA and immunofluorescence technologies. Results We demonstrate that commercially available antibodies can show a significant lack of specificity, and PTM-specific antibodies in particular often recognize non-modified versions of the protein. In addition, detection may be hindered by other PTMs in close vicinity, complicating the interpretation of results. Finally, we compiled a panel of specific antibodies and show that they are useful to detect PTM-modified endogenous tau in hiPSC-derived neurons and mouse brains. Conclusion This study has created a platform to reliably and robustly detect changes in localization and abundance of post-translationally modified tau in health and disease. A web-based version of TauPTM is fully available at http://www.tauptm.org .http://link.springer.com/article/10.1186/s13024-017-0229-1TauPost-translational modificationAntibody validationAlzheimer’s disease |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Ebru Ercan Sameh Eid Christian Weber Alexandra Kowalski Maria Bichmann Annika Behrendt Frank Matthes Sybille Krauss Peter Reinhardt Simone Fulle Dagmar E. Ehrnhoefer |
spellingShingle |
Ebru Ercan Sameh Eid Christian Weber Alexandra Kowalski Maria Bichmann Annika Behrendt Frank Matthes Sybille Krauss Peter Reinhardt Simone Fulle Dagmar E. Ehrnhoefer A validated antibody panel for the characterization of tau post-translational modifications Molecular Neurodegeneration Tau Post-translational modification Antibody validation Alzheimer’s disease |
author_facet |
Ebru Ercan Sameh Eid Christian Weber Alexandra Kowalski Maria Bichmann Annika Behrendt Frank Matthes Sybille Krauss Peter Reinhardt Simone Fulle Dagmar E. Ehrnhoefer |
author_sort |
Ebru Ercan |
title |
A validated antibody panel for the characterization of tau post-translational modifications |
title_short |
A validated antibody panel for the characterization of tau post-translational modifications |
title_full |
A validated antibody panel for the characterization of tau post-translational modifications |
title_fullStr |
A validated antibody panel for the characterization of tau post-translational modifications |
title_full_unstemmed |
A validated antibody panel for the characterization of tau post-translational modifications |
title_sort |
validated antibody panel for the characterization of tau post-translational modifications |
publisher |
BMC |
series |
Molecular Neurodegeneration |
issn |
1750-1326 |
publishDate |
2017-11-01 |
description |
Abstract Background Tau is a microtubule-binding protein, which is subject to various post-translational modifications (PTMs) including phosphorylation, methylation, acetylation, glycosylation, nitration, sumoylation and truncation. Aberrant PTMs such as hyperphosphorylation result in tau aggregation and the formation of neurofibrillary tangles, which are a hallmark of Alzheimer’s disease (AD). In order to study the importance of PTMs on tau function, antibodies raised against specific modification sites are widely used. However, quality control of these antibodies is lacking and their specificity for particular modifications is often unclear. Methods In this study, we first designed an online tool called ‘TauPTM’, which enables the visualization of PTMs and their interactions on human tau. Using TauPTM, we next searched for commercially available antibodies against tau PTMs and characterized their specificity by peptide array, immunoblotting, electrochemiluminescence ELISA and immunofluorescence technologies. Results We demonstrate that commercially available antibodies can show a significant lack of specificity, and PTM-specific antibodies in particular often recognize non-modified versions of the protein. In addition, detection may be hindered by other PTMs in close vicinity, complicating the interpretation of results. Finally, we compiled a panel of specific antibodies and show that they are useful to detect PTM-modified endogenous tau in hiPSC-derived neurons and mouse brains. Conclusion This study has created a platform to reliably and robustly detect changes in localization and abundance of post-translationally modified tau in health and disease. A web-based version of TauPTM is fully available at http://www.tauptm.org . |
topic |
Tau Post-translational modification Antibody validation Alzheimer’s disease |
url |
http://link.springer.com/article/10.1186/s13024-017-0229-1 |
work_keys_str_mv |
AT ebruercan avalidatedantibodypanelforthecharacterizationoftauposttranslationalmodifications AT sameheid avalidatedantibodypanelforthecharacterizationoftauposttranslationalmodifications AT christianweber avalidatedantibodypanelforthecharacterizationoftauposttranslationalmodifications AT alexandrakowalski avalidatedantibodypanelforthecharacterizationoftauposttranslationalmodifications AT mariabichmann avalidatedantibodypanelforthecharacterizationoftauposttranslationalmodifications AT annikabehrendt avalidatedantibodypanelforthecharacterizationoftauposttranslationalmodifications AT frankmatthes avalidatedantibodypanelforthecharacterizationoftauposttranslationalmodifications AT sybillekrauss avalidatedantibodypanelforthecharacterizationoftauposttranslationalmodifications AT peterreinhardt avalidatedantibodypanelforthecharacterizationoftauposttranslationalmodifications AT simonefulle avalidatedantibodypanelforthecharacterizationoftauposttranslationalmodifications AT dagmareehrnhoefer avalidatedantibodypanelforthecharacterizationoftauposttranslationalmodifications AT ebruercan validatedantibodypanelforthecharacterizationoftauposttranslationalmodifications AT sameheid validatedantibodypanelforthecharacterizationoftauposttranslationalmodifications AT christianweber validatedantibodypanelforthecharacterizationoftauposttranslationalmodifications AT alexandrakowalski validatedantibodypanelforthecharacterizationoftauposttranslationalmodifications AT mariabichmann validatedantibodypanelforthecharacterizationoftauposttranslationalmodifications AT annikabehrendt validatedantibodypanelforthecharacterizationoftauposttranslationalmodifications AT frankmatthes validatedantibodypanelforthecharacterizationoftauposttranslationalmodifications AT sybillekrauss validatedantibodypanelforthecharacterizationoftauposttranslationalmodifications AT peterreinhardt validatedantibodypanelforthecharacterizationoftauposttranslationalmodifications AT simonefulle validatedantibodypanelforthecharacterizationoftauposttranslationalmodifications AT dagmareehrnhoefer validatedantibodypanelforthecharacterizationoftauposttranslationalmodifications |
_version_ |
1725239648104480768 |