Detection of Human USP8 by Western Blot.
Western blot shows lysates of U2OS human osteosarcoma cell line and KG‑1 human acute myelogenous leukemia cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human USP8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7735) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for USP8 at approximately 130 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 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.
USP8 (Ubiquitin-Specific-Processing protein 8; also UBPY and ubiquitin carboxyterminal hydrolase 8) is a 125-130 kDa member of the peptidase C19 family of proteins. It is a widely expressed deubiquitinating enzyme that regulates the internalization and trafficking of transmembrane growth factor receptors. Ubiquitinated TKRs are typically targeted for internalization and degradation, and USP8 appears to block this activity. Conversely, USP8 has been shown to have opposite effects on GPCR activity. Here, it accelerates ubiquitinated chemokine receptor (CXCR4) internalization and subsequent inactivation, notably through a deubiquitination-independent mechanism. Human USP8 is 1118 amino acids (aa) in length. It contains one MIT domain that mediates endosomal trafficking (aa 33-116), a rhodanese domain that binds RNF proteins (aa 184-305), an SH3 domain binding sequence (aa 405-413), and a C-terminal catalytic domain that hydrolyses ubiquitin adducts (aa 776‑1106). USP8 is phosphorylated at Ser702, 705, 784 and 787. There is one oncogenic recombination isoform that fuses the p85 beta -subunit of PI3 kinase to the C‑terminal 363 aa of USP8, beginning with Pro746. Full-length human and mouse USP8 share 82% aa sequence identity.
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