Human ECM1 Antibody

Catalog # Availability Size / Price Qty
AF3937
AF3937-SP
Detection of Human ECM‑1 by Western Blot.
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Product Details
Citations (1)
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Human ECM1 Antibody Summary

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 either lyophilized or 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.

Scientific Data

Western Blot Detection of Human ECM-1 antibody by Western Blot. View Larger

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 antibody in Human Colon Cancer Tissue by Immunohistochemistry (IHC-P). View Larger

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.

Simple Western Detection of Human ECM-1 antibody by Simple Western<sup>TM</sup>. View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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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Citation for Human ECM1 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. ECM1 is associated with endocrine resistance in ER+ breast cancers
    Authors: Tae Won Lee, Kyung-min Lee
    Animal Cells and Systems

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