ECM-1/Secretory Component P85 Antibody (966231) [Unconjugated]

Western blot shows lysates of CCD-1070Sk human foreskin fibroblast cell line and WS-1 human fetal skin fibroblast cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human ECM-1 Monoclonal Antibody (Catalog # MAB39371) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). 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 3.

Recombinant human ECM-1 protein was serially diluted 2-fold and captured by Mouse Anti-Human ECM-1 Monoclonal Antibody (Catalog # MAB39371) coated on a Clear Polystyrene Microplate (Catalog # DY990). Mouse Anti-Human ECM-1 Monoclonal Antibody (Catalog # MAB3937) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994).

• Reactivity: Hu
• Applications: WB, ELISA
• Clone: 966231
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

ECM-1/Secretory Component P85 Antibody (966231) [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human ECM-1
  Ala20-Glu540
  Accession # Q16610
• Specificity: Detects human ECM-1 in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • ELISA
  • Western Blot 2 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for ECM-1/Secretory Component P85 Antibody (966231) [Unconjugated]

• ECM1
• extracellular matrix protein 1
• p85
• Secretory Component Glycoprotein
• Secretory Component P85

Background

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(X_{7 ‑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).

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Uniprot:
  • Q16610()