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.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
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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