Detects human HAI-1 in direct ELISAs and Western blots. In direct ELISAs, approximately 20% cross-reactivity with recombinant mouse HAI‑1 is observed and less than 1% cross-reactivity with recombinant human HAI-2 is observed.
Polyclonal Goat IgG
Mouse myeloma cell line NS0-derived recombinant human HAI-1 Pro37-Glu449 Accession # NP_003701
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Detection of Human HAI‑1 by Western Blot.
Western blot shows lysates of MCF‑7 human breast cancer cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human HAI‑1 Ectodomain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1048) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for HAI‑1 at approximately 70 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
HAI‑1 in Human Lung Cancer Tissue.
HAI‑1 was detected in immersion fixed paraffin-embedded sections of human lung cancer tissue using Goat Anti-Human HAI‑1 Ectodomain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1048) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to plasma membrane in epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human HAI‑1 by Simple WesternTM.
Simple Western lane view shows lysates of MCF‑7 human breast cancer cell line, loaded at 0.2 mg/mL. A specific band was detected for HAI‑1 at approximately 88 kDa (as indicated) using 10 µg/mL of Goat Anti-Human HAI‑1 Ectodomain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1048) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system. Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.
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.
HAI-1 is a Kunitz-type serine protease inhibitor, identified as a strong inhibitor of HGF activator (HGFA) and matriptase (1). The membrane-anchored HAI-1 consists of two Kunitz domains, a LDL-receptor-like domain, and a C-terminal transmembrane domain (2). Two soluble forms are generated by ectodomain shedding, one with a single Kunitz domain and the other with two Kunitz domains. HAI-1 is not only an inhibitor but also a specific receptor of active HGFA, acting as a reservoir of this enzyme on the cell surface (3). The shedding of HAI-1 and HGFA/HAI-1 complex is enhanced by treatment with phorbol 12-myristate 13-acetate or IL-1 beta. The regulated shedding is completely inhibited by a synthetic zinc metalloprotease inhibitor (3).
Denda, K. et al. (2002) J. Biol. Chem. 277:14053.
Shimomura, T. et al. (1997) J. Biol. Chem. 272:6370.
Kataoka, H. et al. (2000) J. Biol. Chem. 275:40453.
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