title: A Simple Method to Avoid Nonspecific Signal When Using Monoclonal Anti-Tau Antibodies in Western Blotting of Mouse Brain Proteins.
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.
authors.affiliation: Département de psychiatrie et neurosciences, Université Laval, Québec, QC, Canada.
authors.forename: Franck R
authors.initials: FR
authors.lastname: Petry
authors.affiliation: Department of Neurology, MassGeneral Institute for Neurodegenerative Disease, Alzheimer's Disease Research Laboratory, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA, USA.
authors.forename: Samantha B
authors.initials: SB
authors.lastname: Nicholls
authors.affiliation: Département de psychiatrie et neurosciences, Université Laval, Québec, QC, Canada.|Centre de Recherche du CHU de Québec, Neurosciences, Québec, QC, Canada.
authors.forename: Sébastien S
authors.initials: SS
authors.lastname: Hébert
authors.affiliation: Département de psychiatrie et neurosciences, Université Laval, Québec, QC, Canada. emmanuel@planel.org.|Centre de Recherche du CHU de Québec, Neurosciences, Québec, QC, Canada. emmanuel@planel.org.|Centre Hospitalier de l'Université Laval (CHUL), P0-9800, 2705 Boulevard Laurier, Québec, QC, Canada, G1V 4G2. emmanuel@planel.org.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Planel
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D000918:Antibody Specificity; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism*; D007147:Immunoglobulin Light Chains / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D010766:Phosphorylation; D017354:Point Mutation; D016875:tau Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D007147:Immunoglobulin Light Chains; D016875:tau Proteins
keywords: Alzheimer’s disease; Antibody specificity; Light chain antibody; Mice models; Tau phosphorylation