Human Tenascin C Antibody AF3358: R&D Systems

Human Tenascin C Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human Tenascin C in direct ELISAs and Western blots.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human Tenascin C
    Ser186-Pro625
    Accession # P24821
  • 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
  • Immunocytochemistry
    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 Tenascin C by Western Blot. Western blot shows lysates of U‑87 MG human glioblastoma/astrocytoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Tenascin C Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3358) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for Tenascin C at approximately 250 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
Tenascin C in U‑87 MG Human Cell Line. Tenascin C was detected in immersion fixed U‑87 MG human glioblastoma/astrocytoma cell line using Goat Anti-Human Tenascin C Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3358) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 493-conjugated Anti-Goat IgG Secondary Antibody (green; Catalog # NL003) 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 0.2 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: Tenascin C

Tenascin C, also known as hexabrachion, cytotactin, neuronectin, GMEM, JI, myotendinous antigen, glioma-associated-extracellular matrix antigen, and GP 150‑225, is a member of the Tenascin family of extracellular matrix proteins. It is secreted as a disulfide-linked homohexamer whose subunits can vary in size from approximately 200 kDa to over 300 kDa due to differences in glycosylation (1). Rotary-shadowed electron micrographs of the purified molecule show six strands joined to one another at one end in a globular domain with each arm terminating in a knob-like structure (2, 3). The human Tenascin C monomer is synthesized as a precursor with a 22 amino acid (aa) signal sequence and a 2179 aa mature chain. The mature chain consists of a coiled-coil region (aa 118‑145), followed by 15 EGF‑like domains, 15 fibronectin type-III domains, and a fibrinogen C-terminal domain. In addition, there are 23 potential sites of N-linked glycosylation. Alternative splicing within the fibronectin type-III repeats produces six isoforms for human Tenascin C. Mature human Tenascin C (isoform 1) shares 84% aa sequence identity with mature mouse Tenascin C. In the developing embryo, Tenascin C is expressed during neural, skeletal, and vascular morphogenesis (1, 2). In the adult, it virtually disappears with continued basal expression detectable only in tendon-associated tissues (1, 2). However, great up‑regulation in expression occurs in tissues undergoing remodeling processes seen during wound repair and neovascularization or in pathological states such as inflammation or tumorigenesis (1, 4, 5). Biologically, Tenascin C functions as an adhesion-modulatory extracellular matrix protein (1, 4‑8). Specifically, it antagonizes the adhesive effects of fibronectin, and impacts the ability of fibroblasts to deposit and contract the matrix by affecting the morphology and signaling pathways of adherent cells (5‑7). Tenascin C acts by blocking syndecan-4 binding at the edges of the wound and by suppressing fibronectin-mediated activation of RhoA and focal adhesion kinase (FAK) (4‑8). Tenascin C thus promotes epidermal cell migration and proliferation during wound repair.

  • References:
    1. Hsia, H.C. and J.E. Schwarzbauer (2005) J. Biol. Chem. 280:26641.
    2. Nies, D.E. et al. (1991) J. Biol. Chem. 266:2818.
    3. Erickson, H.P and J.L. Iglesias (1984) Nature 311:267.
    4. Orend, G. et al. (2003) Oncogene 22:3917.
    5. Wenk, M.B. et al. (2000) J. Cell Biol. 150:913.
    6. Midwood, K.S. et al. (2004) Mol. Biol. Cell 15:5670.
    7. Midwood, K.S. and J. E. Schwarzbauer (2002) Mol. Biol. Cell 13:3601.
    8. Hsia, H.C. and J.E. Schwarzbauer (2006) J. Surg. Res. 136:92.
    9.  
  • Entrez Gene IDs:
    3371 (Human); 21923 (Mouse); 116640 (Rat)
  • Alternate Names:
    150-225; Cytotactin; Glioma-associated-extracellular matrix antigen; GMEM; GP 150-225; hexabrachion (tenascin C, cytotactin); hexabrachion (tenascin); Hexabrachion; HXB; HXBcytotactin; JI; MGC167029; Myotendinous antigen; neuronectin; Tenascin C; Tenascin J1; tenascin; tenascin-C isoform 14/AD1/16; Tenascin-C; TNC; TN-C; TNGP
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