Chapter title |
A Simple Method to Avoid Nonspecific Signal When Using Monoclonal Anti-Tau Antibodies in Western Blotting of Mouse Brain Proteins.
|
---|---|
Chapter number | 15 |
Book title |
Tau Protein
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-6598-4_15 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6596-0, 978-1-4939-6598-4
|
Authors |
Franck R. Petry, Samantha B. Nicholls, Sébastien S. Hébert, Emmanuel Planel, Petry, Franck R, Nicholls, Samantha B, Hébert, Sébastien S, Planel, Emmanuel |
Editors |
Caroline Smet-Nocca |
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. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 11 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Researcher | 3 | 27% |
Professor > Associate Professor | 2 | 18% |
Student > Ph. D. Student | 1 | 9% |
Other | 1 | 9% |
Student > Master | 1 | 9% |
Other | 1 | 9% |
Unknown | 2 | 18% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 2 | 18% |
Neuroscience | 2 | 18% |
Immunology and Microbiology | 1 | 9% |
Agricultural and Biological Sciences | 1 | 9% |
Medicine and Dentistry | 1 | 9% |
Other | 1 | 9% |
Unknown | 3 | 27% |