Anti-Tau (3-repeat isoform RD3) antibody | knockout validation | Millipore 05-803

This is a knockout-validated antibody summary, based on the publication "A validated antibody panel for the characterization of tau post-translational modifications", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Anti-Tau (3-repeat isoform RD3) antibody | knockout validation | Millipore 05-803 figure 1
Figure 1. Immunoblotting using hTg-Tau and Tau-KO brain lysates confirms the specificity for tau antibody. From [1].
Antibody information

Mouse monoclonal IgG

Company: Millipore

Antibody: Anti-Tau (3-repeat isoform RD3)

Catalog number: 05-803

Summary: Mouse monoclonal IgG against a synthetic peptide corresponding to amino acids 209-224 of human Tau (isoform RD3). Reacts with human by western blot and immunohistochemistry.

Validation Method

Western blot

Sample

Brains from hTg-Tau and Tau-KO mice. Lysates were prepared by dounce homogenizer in 350 μl of lysis buffer containing 150 mM NaCl, 20 mM Tris pH 7.5, 1 mM EDTA, 1 mM EGTA, 1% Triton-X100 and protease, phosphatase, demethylase (500 μM IOX1, 2 μM Daminozide, 10 μM Paragyline Hydrochloride), deacetylase (10 μM Trichostatin A, 5 mM Nicotinamide), O-GlcNAcase (1 μM Thiamet-G) inhibitors.

Blocking agent

5% BSA in 1X TBST.

Primary incubation

1:500 dilution in blocking solution overnight at 4 °C.

Secondary incubation

1:20,000 dilution IRDye Donkey anti-mouse 800 or IRDye Goat anti-rabbit 680 in blocking solution for 2 h at room temperature.

Detection

Li-Cor Odyssey CLx imaging station.

References
  1. Ercan E, Eid S, Weber C, Kowalski A, Bichmann M, Behrendt A, et al. A validated antibody panel for the characterization of tau post-translational modifications. Mol Neurodegener. 2017;12:87 pubmed publisher