Human ECM1 Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human ECM-1 in direct ELISAs and Western blots. In direct ELISAs, less than 10% cross-reactivity with recombinant mouse ECM‑1 is observed.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human ECM-1
    Ala20-Glu540
    Accession # AAH23505
  • Formulation
    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.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Simple Western
    10 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human ECM‑1 by Western Blot. Western blot shows lysates of COLO 205 human colorectal adenocarcinoma cell line and SK‑Mel‑28 human malignant melanoma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human ECM‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3937) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for ECM‑1 at approximately 75 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunohistochemistry
ECM‑1 in Human Colon Cancer Tissue. ECM‑1 was detected in immersion fixed paraffin-embedded sections of human colon cancer tissue using 15 µg/mL Sheep Anti-Human ECM‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3937) overnight at 4 °C. Tissue was stained with the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific labeling was localized to the plasma membrane of epithelial cells. Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human ECM‑1 by Simple WesternTM. Simple Western lane view shows lysates of COLO 205 human colorectal adenocarcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for ECM‑1 at approximately 90 kDa (as indicated) using 10 µg/mL of Sheep Anti-Human ECM‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3937) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    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.
Background: ECM1

Extracellular matrix protein-1 (ECM-1) is an 85 kDa, secreted glycoprotein important in connective tissue organization (1‑3). Of three identified splice variants the 540 amino acid (aa) form, ECM-1a, is the most widely expressed, with the highest expression in the placenta and heart (2). ECM-1b (415 aa) is found only in tonsil and associated with suprabasal keratinocytes (2, 4). Since ECM-1b expression is differentiation-dependent, a role in terminal keratinocyte differentiation has been suggested (4). ECM-1c (559 aa) accounts for approximately 15% of skin ECM-1 (5). Human ECM-1a contains a 19 aa signal peptide and a 521 aa secreted portion that includes an N-terminal proline-rich, cysteine-free region, two tandem repeat domains, and a C-terminal domain. There are six repeats of a CC(X7 ‑10)C motif (x = any aa) within the tandem repeat and C‑terminal domains. These motifs are involved in ligand binding to members of the albumin family, and are expected to form two (in ECM-1b) or three (in ECM-1a) “double loop” structures (2). Mature human ECM-1a shows 69%, 71%, 72%, and 76% aa identity with corresponding isoforms of mouse, rat, canine, and bovine ECM-1, respectively. ECM-1 is over-expressed in many malignant epithelial tumors and has demonstrated angiogenic activity (6, 7). A variety of ECM-1 mutations, mainly within the first tandem repeat, are considered causative of lipoid proteinosis, a condition showing thickened and irregular extracellular matrix within connective tissue (8). In the autoimmune condition lichen sclerosis, auto-antibodies mainly recognize the second tandem repeat or the C-terminus of ECM-1 (9). These domains also bind the extracellular matrix molecules fibulin-1 and perlecan (5, 10). The phenotypes of lipoid proteinosis and lichen sclerosis support a role for ECM-1 as a “biological glue” in the dermis (1).

  • References:
    1. Chan, I. (2004) Exp. Dermatol. 29:52.
    2. Smits, P. et al. (1997) Genomics 45:487.
    3. Bhalerao, J. et al. (1995) J. Biol. Chem 270:16385.
    4. Smits, P. et al. (2000) J. Invest. Dermatol. 114:718.
    5. Mongiat, M. et al. (2003) J. Biol. Chem. 278:17491.
    6. Han, Z. et al. (2001) FASEB J. 15:988.
    7. Wang, L. et al. (2003) Cancer Lett. 200:57.
    8. Hamada, T. et al. (2003) J. Invest. Dermatol. 120:345.
    9. Oyama, N. et al. (2004) J. Clin. Invest. 113:1550.
    10. Fujimoto, N. et al. (2005) Biochem. Biophys. Res. Commun. 333:1327.
  • Long Name:
    Extracellular Matrix Protein 1
  • Entrez Gene IDs:
    1893 (Human); 13601 (Mouse); 116662 (Rat)
  • Alternate Names:
    ECM1; extracellular matrix protein 1; p85; Secretory Component Glycoprotein; Secretory Component P85
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