Detection of Human UCH‑L3 by Western Blot.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and HT‑29 human colon adenocarcinoma cell line. PVDF membrane was probed with 3 µg/mL of Mouse Anti-Human UCH‑L3 Monoclonal Antibody (Catalog # MAB6008) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for UCH‑L3 at approximately 28 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
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-L3 (Ubiquitin carboxyterminal hydrolase isozyme 3) is a 26‑28 kDa member of the peptidase C12 family of enzymes. It shows wide expression, being noted in adipocytes, renal duct epithelium, neurons, and striated muscle. UCH‑L3 cleaves both monomeric ubiquitin (Ub) from Ub‑protein conjugates, and a GGLRQ peptide from the C‑terminus of the Ub‑like protein NEDD‑8. Notably, UCH-L3 activity is muted in the presence of Ub interacting dimers. Human UCH-L3 is 230 amino acids (aa) in length. It is phosphorylated on Ser75 and Ser130 and contains two ubiquitin-binding sequences between aa 40‑57 and 178‑186. Four potential splice forms have been reported. Two show a four aa deletion between aa 15‑18, with one also containing a 14 aa substitution for aa 179‑230. Two others contain an alternative start site at Met37, one of which is also accompanied by a seven aa substitution for aa 143‑230. Full length human UCH-L3 shares 98% aa identity with mouse and rat UCH‑L3.
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