Immunohistochemistry was performed on 5-µm-thick sections of formalin-fixed, paraffin-embedded tissue. Glass-mounted sections were deparaffinized in xylene and rehydrated in ethanol and distilled water. Immunohistochemistry for tau used an antibody to phospho-serine 202 (CP13, mouse monoclonal; from Peter Davies, PhD, Feinstein Institute, North Shore Hospital, NY). Nine sections, which covered almost all major anatomical regions affected in CBD, were processed for immunohistochemistry for phospho-TDP-43 (pS409/410, mouse monoclonal, 1:5,000; Cosmo Bio, Tokyo, Japan) according to previously published methods.21 (link)All immunohistochemistry was performed using a DAKO AutostainerPlus (Agilent/DAKO, Santa Clara, CA) with the DAKO EnVision + system-HRP with 3,3′-diaminobenzidine (DAB) as the chromogen. Nonspecific antibody binding was blocked with normal goat serum (Sigma, St. Louis, MO).
Protocol detail
Immunohistochemistry of Tau and TDP-43 in CBD
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Corresponding Organization :
Other organizations : University of California, San Diego, Mayo Clinic in Florida, Mount Sinai Medical Center, Mayo Clinic, Mayo Clinic in Arizona
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Variable analysis
independent variables
- Antibody to phospho-serine 202 (CP13, mouse monoclonal; from Peter Davies, PhD, Feinstein Institute, North Shore Hospital, NY)
- Antibody to phospho-TDP-43 (pS409/410, mouse monoclonal, 1:5,000; Cosmo Bio, Tokyo, Japan)
- Immunohistochemistry for tau
- Immunohistochemistry for phospho-TDP-43
- 5-µm-thick sections of formalin-fixed, paraffin-embedded tissue
- Glass-mounted sections
- Deparaffinization in xylene and rehydration in ethanol and distilled water
- DAKO AutostainerPlus (Agilent/DAKO, Santa Clara, CA) with the DAKO EnVision + system-HRP with 3,3′-diaminobenzidine (DAB) as the chromogen
- Blocking of nonspecific antibody binding with normal goat serum (Sigma, St. Louis, MO)
- Positive control: Nine sections covering almost all major anatomical regions affected in CBD
- Negative control: Not explicitly mentioned
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