Immunohistochemistry was performed as previously described.23 (link) Paraffin-embedded tissues were sectioned in a coronal plane at 10–20 μm. The tissue sections were rehydrated, blocked with blocking solution (1% H2O2), and incubated with p-Tau (S202/T205) (1:200), PP2B/PPP3CA (1:200; SantaCruz Biotech), p-Tau (S199) (1:200; Abcam) and anti-III tubulin antibody (1:500 dilutions; Sigma) for 24 h. After washing three times, the slides were processed with Vector ABC Kit (Vector Lab). The immunoreactive signals were developed with DAB chromogen (Thermo Fisher Scientific, Meridian, Rockford, IL, USA) and analyzed under a bright field microscope.
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Immunohistochemical Analysis of Tau Phosphorylation
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Anti antibody
Chromogen
Dilutions
H2o2
Immunohistochemistry
Meridian
Microscope
Paraffin
Ppp3ca
Tissue
Tubulin
Vector
Corresponding Organization : Seoul National University Hospital
Other organizations : Korea Institute of Brain Science, Korea Institute of Science and Technology, Macrogen (South Korea), Seoul National University, Boston University, VA Boston Healthcare System
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Variable analysis
independent variables
- Antibody dilution: p-Tau (S202/T205) (1:200), PP2B/PPP3CA (1:200), p-Tau (S199) (1:200), anti-III tubulin (1:500)
- Immunoreactive signals developed with DAB chromogen and analyzed under a bright field microscope
- Paraffin-embedded tissue sections were rehydrated, blocked with blocking solution (1% H2O2), and incubated with the antibodies for 24 h
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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