|Detection of Human ADAMTS8 by Western Blot. Western blot shows lysates of 293T human embryonic kidney cell line either transfected with human ADAMTS8, mock transfected (-), or transfected with an unrelated vector. PVDF Membrane was probed with 1 µg/mL of Human ADAMTS8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6614) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for ADAMTS8 at approximately 80-85 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.|
|ADAMTS8 in Human Brain. ADAMTS8 was detected in immersion fixed paraffin-embedded sections of human brain (cerebellum) using Sheep Anti-Human ADAMTS8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6614) at 3 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-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to Purkinje neurons. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.|
ADAMTS8 (A disintegrin and metalloprotease with thrombospondin motifs 8; also METH-2) is a 95 kDa member of an ADAMTS subfamily of Zn metalloproteases that includes ADAMST-1, -4, -5 and -15. It is expressed by chrondrocytes, neurons, astrocytes and macrophages, and likely participates in proteoglycan (aggrecan) proteolysis. Human proADAMTS8 is a secreted, 863 amino acid (aa) glycoprotein. It is highly modular and contains a proregion (aa 27-213), a peptidase M12B domain (aa 219-429), a disintegrin region (aa 438-525), and two TSP type I sequences (aa 526-888) that are separated by an intervening spacer domain (aa 690-831). Cleavage of the proregion generates a mature 80 kDa molecule that may undergo additional processing to create a 65-67 kDa truncated form. There are two potential splice variants. One shows a 31 aa substitution for aa 411-889, while another shows an alternative start site at Met231. Over aa 29-691, human ADAMTS8 shares 79% aa identity with mouse ADAMTS8.
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