Detection of Human SHARPIN by Western Blot.
Western blot shows lysates of U2OS human osteosarcoma cell line and THP‑1 human acute monocytic leukemia cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human SHARPIN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8100) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for SHARPIN at approximately 43 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
SHARPIN in Human Ovary.
SHARPIN was detected in immersion fixed paraffin-embedded sections of human ovary using Sheep Anti-Human SHARPIN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8100) at 10 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
Reconstitute at 0.2 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.
Sharpin (Shank-associated RH domain-interacting protein), also called SIPL1 (Shank interacting protein-like 1) is a 40-45kDa protein. It is an essential component of the cytosolic LUBAC ubiquitin conjugation complex. It conjugates linear ubiquitin chains to IKK gamma and activates NFkB, thus regulating immune and inflammatory responses. Sharpin is widely expressed, with upregulated expression in multiple tumor types. Human Sharpin is a 387 amino acid (aa) protein containing self association, SHANK1 interaction, and zinc finger domains. A 326 aa isoform diverges after aa 308 and lacks the zinc finger domain. Within aa 27-182, human Sharpin shares 67 and 66% aa sequence identity with mouse and rat Sharpin, respectively.
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