Human COMP/Thrombospondin-5 Antibody AF3134: R&D Systems

Human COMP/Thrombospondin-5 Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human COMP in direct ELISAs and Western blots. In Western blots, less than 1% cross-reactivity with recombinant human Thrombospondin-1, -2, -3 and -4 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human COMP/Thrombospondin-5
    Gln21-Ala757 (Ala256Arg)
    Accession # P49747
  • 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.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.05 µg/mL
    See below
  • Neutralization
    Measured by its ability to neutralize COMP/Thrombospondin‑5-mediated adhesion of the ATDC5 mouse chondrogenic cell line. The Neutralization Dose (ND50) is typically 1-4 µg/mL in the presence of 10 µg/mL Recombinant Human COMP/Thrombospondin‑5.
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 COMP/Thrombospondin‑5 by Western Blot. Western blot shows lysates of human cartilage tissue. PVDF membrane was probed with 0.05 µg/mL of Goat Anti-Human COMP/Thrombospondin‑5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3134) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for COMP/Thrombospondin‑5 at approximately 90 and 110 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Cell Adhesion Mediated by COMP/Thrombospondin‑5 and Neutralization by Human COMP/Thrombospondin‑5 Antibody. Recombinant Human COMP/Thrombospondin‑5 (Catalog # 3134-CP), immobilized onto a microplate, supports the adhesion of the ATDC5 mouse chondrogenic cell line in a dose-dependent manner (orange line). Adhesion elicited by Recombinant Human COMP/Thrombospondin‑5 (10 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human COMP/Thrombo­spondin‑5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3134). The ND50 is typically
1-4 µg/mL.
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: COMP/Thrombospondin-5

Cartilage Oligomeric Matrix Protein (COMP), also known as Thrombospondin-5, is a 110 kDa multidomain calcium binding protein that associates with other extracellular matrix molecules. Thrombospondin-1 and -2 constitute subgroup A and form homotrimers, whereas Thrombospondin-3, -4, and COMP constitute subgroup B and form homopentamers (1-4). The human COMP cDNA encodes a 757 amino acid (aa) precursor that includes a 20 aa signal sequence followed by a non-collagenous
coiled‑coil domain, four EGF-like repeats, seven TSP type-3 repeats, and a globular TSP C-terminal domain (5). Human COMP shares 86-93% aa sequence identity with rat, mouse, equine, bovine, and canine COMP. Within the TSP type-3 repeats and TSP C-terminal domain, human COMP shares 60%, 61%, 74%, and 80% aa sequence identity with human Thrombospondin-1, -2, -3, and -4, respectively. The coiled coil domain mediates the association of COMP into disulfide-linked homopentamers with a central hub and peripheral globular domains connected by flexible strands (6, 7). An axial pore is formed by the coiled coil assembly and binds vitamin D3 which is involved in bone and cartilage metabolism (8). An RGD sequence in the third TSP type-3 repeat mediates chondrocyte attachment via Integrin alpha 5 beta 1, although when reduced and in the absence of calcium, attachment is mediated via Integrin alpha V beta 3 (9). COMP is upregulated in rheumatoid arthritis and osteoarthritis, hepatocellular carcinomas, chronic pancreatitis, and pancreatic carcinomas (10-12). Elevated circulating COMP levels are used as a biomarker for early onset of some skeletal disorders (10). Several mutations are associated with skeletal dysplasias, and the most common, a point mutation in the third TSP type-3 repeat, results in diminished calcium binding ability (13, 14).

  • References:
    1. Adams, J.C. and J. Lawler (2004) Int J. Biochem. Cell Biol. 36:961.
    2. Posey, K.L. et al. (2004) Int. J. Biochem. Cell Biol. 36:1005.
    3. Adams, J.C. (2004) Int. J. Biochem. Cell Biol. 36:1102.
    4. Mann, H.H. et al. (2004) J. Biol. Chem. 279:25294.
    5. Newton, G. et al. (1994) Genomics, 24:435.
    6. DiCesare, P. et al. (1995) J. Orthopaedic Res. 13:422.
    7. Efimov, V.P. et al. (1994) FEBS Lett. 341:54.
    8. Ozbek, S. et al. (2002) EMBO J. 21:5960.
    9. Chen, F.H. et al. (2005) J. Biol. Chem. 280:32655.
    10. Wislowska, M. and B. Jablonska (2005) Clin. Rheumatol. 24:278.
    11. Xiao, Y. et al. (2004) J. Gastroenterol. Hepatol. 19:296.
    12. Liao, Q. et al. (2003) Scand. J. Gastroenterol. 38:207.
    13. Kennedy, J. et al. (2005) Eur. J. Hum. Genet. 13:547.
    14. Hou, J. et al. (2000) Cell Calcium 27:309.
  • Long Name:
    Cartilage Oligomeric Matrix Protein
  • Entrez Gene IDs:
    1311 (Human); 12845 (Mouse)
  • Alternate Names:
    cartilage oligomeric matrix protein (pseudoachondroplasia, epiphyseal dysplasia1, multiple); cartilage oligomeric matrix protein; cartilage oligomeric matrix protein(pseudoachondroplasia, epiphyseal dysplasia1, multiple); COMP; EDM1; EPD1; MED; MEDMGC131819; MGC149768; PSACH; pseudoachondroplasia (epiphyseal dysplasia 1, multiple); THBS5; Thrombospondin5; Thrombospondin-5; TSP5
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