|Detection of Human Thrombospondin‑1 by Western Blot. Western blot shows lysates of MDA‑MB‑231 human breast cancer cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Thrombospondin‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3074) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for Thrombospondin‑1 at approximately 160 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
|Thrombospondin‑1 in Human Colon. Thrombospondin‑1 was detected in immersion fixed paraffin-embedded sections of human colon using Goat Anti-Human Thrombospondin‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3074) at 1.7 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 the cytoplasm of epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.|
Detection of Human Thrombospondin‑1 by Simple WesternTM. Simple Western lane view shows lysates of MBA‑MB‑468 human breast cancer cell line, loaded at 0.2 mg/mL. A specific band was detected for Thrombospondin‑1 at approximately 221 kDa (as indicated) using 10 µg/mL of Goat Anti-Human Thrombospondin‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3074) 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.
Thrombospondin-1 (TSP-1) is a 150‑180 kDa charter member of the thrombospondin family of extracellular matrix proteins. Human TSP‑1 is synthesized as an 1170 amino acid (aa) precursor that contains an 18 aa signal sequence and 1152 aa mature molecule. The mature molecule has been described as containing three distinct regions that create the shape of a dumbbell. There is an intital, 140 aa N-terminal laminin G-like globular region that binds heparin (aa 19‑258). This is followed by an extended, central collagen-binding region that contains one type C von Willebrand factor domain, plus three TSP type I and three TSP type II (or EGF-like) domains (aa 259‑712). The C‑terminus (aa 713‑1170) appears as a large globule with two halves; one calcium‑binding region (aa 713‑950) with seven Asp‑rich TSP type III domains, and one terminal region (aa 951‑1170) with TSP‑unique motifs (1). This C-terminal region is believed to mediate CD47 and cell binding (2‑5). The TSP type I repeats have multiple functions. They bind to type V collagen, laminin, fibronectin and CD36. They also contain a recognition site for C‑mannosylation on Trp. Finally, a type I KRFK motif induces the release of mature TGF-b from LAP. This is an effect not found in TSP-2. The function of the type II repeats in unclear. TSP‑1 is secreted as a disulfide-linked 450 kDa homotrimer. The cysteines responsible lie just N‑terminal to the first type I TSP repeat. Mature human TSP-1 is 61% aa identical to human TSP-2. It is also 95%, 97% and 95% aa identical to mouse, dog and rat TSP-1, respectively.
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Western Blot: Human Thrombospondin-1 Antibody [AF3074]