This is a knockout-validated antibody summary, based on the publication "Tau-positive nuclear indentations in P301S tauopathy mice", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Company: Calbiochem

Antibody: tau

Catalog number: 577801

Summary: This Anti-Tau Mouse mAb (TAU-5) is validated for use in Frozen Sections, Immunoblotting, Immunoprecipitation, Paraffin Sections for the detection of Tau. Recognizes the ~45-68 kDa Tau protein.

IHC, immunohistochemistry.

Mouse brains were immediately removed and dissected on an ice-cold plate and left hemispheres, processed for histology, were placed in 4% paraformaldehyde in Sorensen’s phosphate buffer overnight and then immersed in 30% sucrose in PBS for 72 h. Once cryoprotected, the samples were included in OCT compound (Sakura Finetek Europe), frozen and stored at -80°C until use. 30 μm sagittal sections were cut on a cryostat (Thermo Scientific), collected and stored free floating in glycol containing buffer (30% glycerol, 30% ethylene glycol in 0,02M phosphate buffer) at -20°C. Sections were immersed in 0.3% H2O2 in PBS for 30 min to quench endogenous peroxidase activity.

Sections were blocked for 1 h in PBS containing 0.5% Fetal Bovine Serum, 0.3% Triton X-100 and 1% BSA (Sigma-Aldrich).

Incubated overnight at 4°C in PBS containing 0.3% Triton X-100 and 1% BSA with the corresponding primary antibody.

Brain sections were incubated with anti-mouse secondary antibody and avidin-biotin complex using the Elite Vectastain kit (Vector Laboratories). Chromogen reactions were performed with diaminobenzidine (SIGMAFASTTM DAB, Sigma) for 10 min.

Mouse sections were mounted on glass slides and coverslipped with Mowiol (Calbiochem). Images were captured using an Olympus BX41 microscope with an Olympus camera DP-70 (Olympus Denmark A/S).

See Fig 2 in [1]. Cortical images of Tau5 immunohistochemistry on sagittal sections of WT mice. Tau5 immunohistochemistry on Tau KO mice was performedas a negative control.