|Detection of Human MMP‑3 by Western Blot. Western blot shows Recombinant Human MMP-3 Westrern Blot Standard Protein (2 μL, Catalog # WBC015). PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human MMP‑3 Monoclonal Antibody (Catalog # MAB513) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for MMP‑3 at approximately 55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
|MMP‑3 in Human Prostate and Human Prostate Cancer Tissue. MMP‑3 was detected in immersion fixed paraffin-embedded sections of normal human prostate and human prostate cancer tissue using Mouse Anti-Human MMP‑3 Monoclonal Antibody (Catalog # MAB513) at 15 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 cytoplasm in cancer cells (right panel). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.|
|MMP‑3 in MG-63 Human Cell Line. MMP‑3 was detected in immersion fixed MG-63 human osteosarcoma cell line using 10 µg/mL Mouse Anti-Human MMP‑3 Monoclonal Antibody (Catalog # MAB513) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.|
Intracellular Staining by Flow Cytometry
|Detection of MMP‑3 in MG‑63 Human Cell Line by Flow Cytometry. MG‑63 human osteosarcoma cell line was stained with Mouse Anti-Human MMP‑3 Monoclonal Antibody (Catalog # MAB513, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.|
Matrix metalloproteinases are a family of zinc and calcium dependent endopeptidases with the combined ability to degrade all the components of the extracellular matrix. MMP-3 (stromelysin-1), can degrade a broad range of substrates including collagen alpha chains, aggrecan, laminin, fibronectin, elastin, casein, alpha -1 antitrypsin, myelin basic protein, IL-1 beta, IGFBP-3, pro MMP-1, pro MMP-7, pro MMP-8, pro MMP-9 and pro MMP-13. MMP-3 does not cleave the triple helical region of interstitial collagens, a characteristic which distinguishes the stromelysins from the collagenases. The MMP-3 substrate repertoire extends beyond extracellular matrix proteins and implicates MMP-3 in roles other than direct tissue remodelling, for instance, enzyme cascades and cytokine regulation. MMP-3 is expressed by fibroblasts, chrondrocytes, osteoblasts, endothelial cells, smooth muscle cells and macrophages. Structurally, MMP-3 may be divided into several distinct domains; a pro-domain which is cleaved upon activation; a catalytic domain containing the zinc binding site; a short hinge region and a carboxyl terminal (hemopexin-like) domain.
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