Tau antibody | knockout validation | Invitrogen MN1050
DOI
//dx.doi.org/10.13070/ko.en.7.2230
Date
2017-04-08

This is a knockout-validated antibody summary, based on the publication "Specificity of anti-tau antibodies when analyzing mice models of Alzheimer's disease: problems and solutions", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Tau antibody | knockout validation | Invitrogen MN1050 figure 1
Figure 1. Western blot of Tau-1 in the cortex of 3 mouse lines: control mice (WT and Hypothermic), Tau KO mice and 3xTg mice, with n = 3 for each group. Normal anti-mouse (1) and mouse TrueBlot ULTRA (2) secondary antibodies were used to detect primary antibodies. The heat stable fraction was used to remove non-specificity (3). From [1].
Antibody information

Mouse monoclonal IgG1, kappa

Company: Invitrogen

Antibody: Tau

Catalog number: MN1050

Summary: Mouse monoclonal IgG1, kappa against purified denatured bovine microtubule associated proteins. Reacts with human, and has been published to react also with rat, primate, fruit fly, hamster, mouse, and chicken. Suitable for western blot, immunohistochemistry and immunofluorescence. Published also to work in ELISA and immunoprecipitation.

Validation Method

Western blot

Sample

Mouse brain cortex from TKO, 3xTg-AD, WT and hypothermic mice. Frozen cortices were homogenized in cold radioimmunoassay precipitation (RIPA) buffer (50 mM Tris-HCl, pH 7,4, 1% NP-40, 150 mM NaCl, 0,25% Na-Deoxycholate, 1 mM EDTA, 1 mM Na3VO4, 1 mM NaF, 1 mM PMSF and 10 ml/ml of proteases inhibitors P8340, Sigma Aldrich, Saint-Louis, MO, USA).

Blocking agent

5% dry milk for 1 h at room temperature.

Primary incubation

1/1,000 dilution overnight.

Secondary incubation

1/5,000 dilution goat anti-mouse Ig HRP (Jackson Immunoresearch Laboratories, 115-035-003) for 1h or 1/1,000 dilution TrueBlot ULTRA anti-mouse Ig HRP (eBiosciences, 18-8817-33) overnight .

Detection

ECL (Enhanced Chemiluminescence) detection kit (Millipore, Billerica, MA, USA) and visualized with a Fujifilm LAS4000 imaging system (Fujifilm Life Science USA, Stamford, CT).

Note

Antibody shows moderate unspecificity. Normal (HC+LC) anti-mouse Ig secondary antibodies may be used and TrueBlot or anti-LC secondary antibodies give better tau signal.

References
  1. Petry F, Pelletier J, Bretteville A, Morin F, Calon F, Hébert S, et al. Specificity of anti-tau antibodies when analyzing mice models of Alzheimer's disease: problems and solutions. PLoS ONE. 2014;9:e94251 pubmed publisher