Human ADAMTS4 Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human ADAMTS4 in direct ELISAs and Western blots. In direct ELISAs and Western blots, no cross-reactivity with recombinant human (rh) ADAMTS1, rhADAMTS5, rhADAMTS13, or rhADAMTSL1.2 is observed.
  • Source
    Monoclonal Mouse IgG2A Clone # 416610
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human ADAMTS4
    Phe213-Cys685
    Accession # O75173
  • 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
    Recombinant Human ADAMTS4 (Catalog # 4307-AD) under non-reducing conditions only
  • Immunoprecipitation
    25 µg/mL
    Conditioned cell culture medium spiked with Recombinant Human ADAMTS4 (Catalog # 4307-AD), see our available Western blot detection antibodies
  • Immunocytochemistry
    8-25 µg/mL
    See below
  • Neutralization
    Measured by its ability to neutralize Recombinant Human ADAMTS4 (5 µg/mL, Catalog # 4307-AD) cleavage of the substrate Recombinant Human Aggrecan G1-IGD-G2 Domain (100 µg/mL, Catalog # 1220-PG). The Neutralization Dose (ND50) is typically 25 µg/mL.
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
Immunocytochemistry
ADAMTS4 in SH‑SY5Y Human Cell Line. ADAMTS4 was detected in immersion fixed SH‑SY5Y human neuroblastoma cell line using Mouse Anti-Human ADAMTS4 Monoclonal Antibody (Catalog # MAB4307) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 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: ADAMTS4

ADAMTS4 (a disintegrin and metalloproteinase with thrombospondin motifs 4), also known as aggrecanase-1, is a member of the family of secreted zinc proteases with a multi-domain structure (1‑3). The protein precursors consist of a signal peptide and the following domains: pro, catalytic, disintegrin-like, TS type 1 motif, cysteine-rich, and spacer. It is the only ADAMTS identified that has one TS type I motif. It is an active protease effectively cleaving alpha -2-macroglobulin and aggrecan at multiple sites, and is inhibited by TIMP-3 with inhibition constants in subnanomolar range (4‑6). It receives great attention due to the elevation in its mRNA level after treatment with Interleukin-1 (7). However, in a mouse model of osteoarthritis, ADAMTS4 knock-out mice did not exhibit any significant protective effect (8). ADAMTS4 consists of a signal peptide (aa 1 to 51), a pro domain (aa 52 to 212) and a mature chain (aa 213 to 837) containing the following domains: catalytic (aa 218 to 425), disintegrin (aa 437 to 519), TSP type-1 (aa 520 to 575), Cys-rich (aa 577 to 685) and spacer (aa 686 to 837). The amino acid sequence of rhADAMTS4 is 100%, 97% and 94% identical to that of chimpanzee, dog, and mouse/rat/bovine.

  • References:
    1. Totorella, M. D. et al. (1999) Science 284:1664.
    2. Porter, S. et al. (2005) Biochem. J. 386:15.
    3. Nagase, H. and M. Kashiwagi (2003) Arthritis Res. Ther. 5:94.
    4. Tortorella, M. D. et al. (2004) J. Biol. Chem. 279:17554.
    5. Struglics, A. et al. (2006) Osteoarthritis Cartilage. 14:101.
    6. Kashiwagi, M. et al. (2001) J. Biol. Chem. 276:12501.
    7. Pratta, M. A. et al. (2003) Arthritis Reum. 48:119.
    8. Glasson, S. S. et al. (2004) Arthritis Reum. 50:2547.
  • Long Name:
    A Disintegrin-like and Metalloproteinase Domain with Thrombospondin Motifs 4
  • Entrez Gene IDs:
    9507 (Human); 66015 (Rat)
  • Alternate Names:
    a disintegrin-like and metalloprotease (reprolysin type) with thrombospondintype 1 motif, 4; ADAM metallopeptidase with thrombospondin type 1 motif, 4; ADAM-TS 4; ADAMTS2; ADAMTS-2; ADAMTS4; ADAM-TS4; ADAMTS-4; ADMP-1; ADMP-1EC 3.4.24.82; Aggrecanase 1; aggrecanase-1; EC 3.4.24; KIAA0688A disintegrin and metalloproteinase with thrombospondin motifs 4
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