Detection of Human Ubiquitin+1 by Western Blot.
Western blot shows samples of Recombinant Human Ubiquitin (Catalog # 701‑UB) (2 ng) and Recombinant Human Ubiquitin+1 (Catalog # 703‑UB) (2, 1, and 0.5 ng). PVDF membrane was probed with 1 µg/mL Rabbit Anti-Human Ubiquitin+1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF703) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band for Ubiquitin+1 was detected at approximately 13 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
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 (1, 2).
van Leeuwen, F.W. et al. (1998) Science 279:242.
van Leeuwen, F.W. et al. (1998) Trends Neurosci. 21:331.
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