Abstract
In Alzheimer’s disease and other tauopathies, tau displays several abnormal post-translation modifications such as hyperphosphorylation, truncation, conformation, and oligomerization. Mouse monoclonal antibodies have been raised against such tau modifications for research, diagnostic, and therapeutic purposes. However, many of these primary antibodies are at risk of giving nonspecific signals in common Western blotting procedures. Not because they are unspecific, but because the secondary antibodies used to detect them will also detect the heavy chain of endogenous mouse immunoglobulins (Igs), and give a nonspecific signal at the same molecular weight than tau protein (around 50 kDa). Here, we propose the use of anti-light chain secondary antibodies as a simple and efficient technique to prevent nonspecific Igs signals at around 50 kDa. We demonstrate the efficacy of this method by removing artifactual signals when using monoclonal antibodies directed at tau phosphorylation (AT100, 12E8, AT270), tau truncation (TauC3), tau oligomerization (TOMA), or tau abnormal conformation (Alz50), in wild-type, 3×Tg-AD, and tau knockout mice.
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Acknowledgments
We would like to thank Dr. Peter Davies (Albert Einstein College of Medecine, Bronx, NY, USA) for the gift of the Alz50 antibody , Dr. Rakez Kayed (Mitchell Center for Neurodegenerative Diseases, Texas, USA) for the TOMA antibody, and Dr. Peter Seubert (Neotope Biosciences, CA, USA) for the gift of the 12E8 antibody.
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Petry, F.R., Nicholls, S.B., Hébert, S.S., Planel, E. (2017). A Simple Method to Avoid Nonspecific Signal When Using Monoclonal Anti-Tau Antibodies in Western Blotting of Mouse Brain Proteins. In: Smet-Nocca, C. (eds) Tau Protein. Methods in Molecular Biology, vol 1523. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6598-4_15
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DOI: https://doi.org/10.1007/978-1-4939-6598-4_15
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