Detection of Human UCH-L1/PGP9.5 by Western Blot.
Western blot shows lysates of human brain (cortex) tissue. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human UCH-L1/PGP9.5 Monoclonal Antibody (Catalog # MAB6007) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for UCH-L1/PGP9.5 at approximately 29 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
UCH-L1/PGP9.5 in Human Prostate.
UCH-L1/PGP9.5 was detected in immersion fixed paraffin-embedded sections of human prostate using Mouse Anti-Human UCH-L1/PGP9.5 Monoclonal Antibody (Catalog # MAB6007) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm of glandular epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human and Mouse UCH-L1/PGP9.5 by Simple WesternTM.
Simple Western lane view shows lysates of human brain (cortex) tissue and mouse brain (cortex) tissue, loaded at 0.2 mg/mL. A specific band was detected for UCH-L1/PGP9.5 at approximately 32 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human UCH-L1/PGP9.5 Monoclonal Antibody (Catalog # MAB6007). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
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.
UCH-L1 (ubiquitin carboxyterminal hydrolase isozyme 1; also PGP9.5) is a 24-27 kDa member of the peptidase C12 family of enzymes. It shows restricted expression, being found in neurons and oocytes. UCH-L1 has dual enzymatic activity. As a monomer, it is a ubiquitin hydrolase that removes ubiquitin from modified proteins; as a homodimer, it acts as a ligase that creates ubiquitin dimers. In neurons, UCH-L1’s most important role appears to be that of generating free ubiquitin. Human UCH-L1 is 223 amino acids (aa) in length. It is O-glycosylated, ubiquitinated, and farnesylated; when farnesylated, it becomes associated with cell membranes. Three potential splice forms are reported. One shows a two aa substitution for aa 12-15, a second contains an alternative start site at Met82, and a third shows the same start site coupled with a deletion of aa 138-153. Full-length human UCH-L1 shares 95% aa identity with mouse UCH-L1.
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