Ubiquitin/Ubiquitin+1 in Human Alzheimer's Disease Brain. Ubiquitin/Ubiquitin+1 was detected in immersion fixed paraffin-embedded sections of human Alzheimer's disease brain (cortex) using 25 µg/mL Mouse Anti-Human Ubiquitin/|
Ubiquitin+1 Monoclonal Antibody (Catalog # MAB701) overnight at 4 °C. Tissue was stained with the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific labeling was localized to the cytoplasm of neurons in the cortex. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
|Detection of Human Ubiquitin/Ubiquitin+1 by Western Blot. Western blot shows samples of Recombinant Human Ubiquitin (Catalog # 701-UB) (2, 1, and 0.5 ng) and Recombinant Human Ubiquitin+1 (Catalog # 703-UB) (2, 1, and 0.5 ng). PVDF membrane was probed with 1-2 µg/mL Mouse Anti-Human Ubiquitin/Ubiquitin+1 Monoclonal Antibody (Catalog # MAB701) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). Specific bands for Ubiquitin and Ubiquitin+1 were detected at approximately 11 kDa and 13 kDa, respectively (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 9.|
Ubiquitin+1 has a carboxyl terminal amino acid sequence that differs from normal Ubiquitin. The different carboxyl terminal sequence appears to result from a frameshift in the Ubiquitin mRNA. The underlying mechanisms creating the mRNA frameshift are not clearly understood. The occurrence of the frameshift that generates Ubiquitin+1 is much more prevalent in patients with Alzheimers Disease or with Down Syndrome than in control individuals who are not afflicted with the disorders. The monoclonal anti-Ubiquitin+1 (Catalog # MAB703) and rabbit polyclonal anti-Ubiquitin+1 (Catalog # AF703) antibodies were raised against the Ubiquitin+1 carboxyl terminal sequence that differs from normal Ubiquitin and are therefore non-reactive with Ubiquitin. Monoclonal anti-Ubiquitin (Catalog # MAB701) detects both Ubiquitin and Ubiquitin+1 indicating that the epitope recognized by this antibody is contained in the portion of the proteins that are identical.
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