Human OX40/TNFRSF4 Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human OX40/TNFRSF4 in direct ELISAs and Western blots. In direct ELISAs and Western blots, less than 1% cross-reactivity with recombinant mouse OX40 is observed.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human OX40
    Leu29-Ala216
    Accession # P43489
  • 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
    0.5 µg/mL
    See below
  • Flow Cytometry
    0.25 µg/106 cells
    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 OX40/TNFRSF4 by Western Blot. Western blot shows lysates of human T regulatory cells (Tregs) and human peripheral blood mononuclear cells (PBMCs) untreated (-) or treated (+) with 1 μg/mL PHA for 5 days. PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human OX40/TNFRSF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3388) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for OX40/TNFRSF4 at approximately 45 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of OX40/TNFRSF4 in Human T Cells by Flow Cytometry. Human T cells were treated for 3‑5 days with 1 μg/mL PHA then stained with Sheep Anti-Human OX40/
TNFRSF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3388, filled histogram) or control antibody (Catalog # 5-001-A, open histogram), followed by NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # NL010).
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: OX40/TNFRSF4

OX40 (CD134; TNFRSF4) is a T cell costimulatory molecule of the TNF receptor superfamily that coordinates with other membrane-bound costimulators such as CD28, CD40, CD30, CD27 and 4-1BB (1‑3). OX40 is expressed on naïve CD4+ T cells only after engagement of the TCR by antigen presenting cells (APC; dendritic and B cells), and costimulation by CD40/CD40 ligand and CD28/B7. It is maximal at 2‑5 days post activation, or 4 hours post reactivation of memory T cells (3‑6). Human OX40 is a 48 kDa type I transmembrane glycoprotein with a 28 amino acid (aa) signal sequence, a 185 aa extracellular domain (ECD) that has four TNFR-Cys repeats and an O-glycosylated hinge region, a 20 aa transmembrane segment, and a 41 aa cytoplasmic domain (3). The ECD of human OX40 shows 71%, 68%, 67%, 64% and 64% aa identity with feline, canine, rabbit, mouse and rat OX40 ECD, respectively. Engagement of OX40 on activated CD4+ T cells by OX40 ligand on activated dendritic cells promotes T cell survival and proliferation, prolongs the immune response, and enhances the number of cells making the transition from effector to memory T cells (1‑6). OX40 signal transduction includes binding TNF receptor-associated factors (TRAFs), and activating NF kappa B and PI3 kinase to enhance expression of cytokines, antiapoptotic Bcl-2 family members, survivin and the chemokine receptor CXCR5 (5‑8). CXCR5 promotes T cell migration to germinal centers to deliver B cell help (5). Studies using knockout or transgenic mice, and agonistic or blocking antibodies, show that OX40/OX40L interaction is critical for establishing or reactivating memory T cells and breaking immune tolerance (9). Blockade of OX40 engagement is efficacious in animal models of allergic airway inflammation, graft-versus-host disease and autoimmune disease (10‑14).

 

  • References:
    1. Salek-Ardakani, S. and M. Croft (2006) Vaccine 24:872.
    2. Hori, T. (2006) Int. J. Hematol. 83:17.
    3. Latza, U. et al. (1994) Eur. J. Immunol. 24:677.
    4. Murata, K. et al. (2000) J. Exp. Med. 191:365.
    5. Fillatreau, S and D. Gray (2003) J. Exp. Med. 197:195.
    6. Gramaglia, I. et al. (1998) J. Immunol. 161:6510.
    7. Rogers, P.R. et al. (2001) Immunity 15:445.
    8. Song, J. et al. (2005) Immunity 22:621.
    9. Bansal-Pakala, P. et al. (2001) Nat. Med. 7:907.
    10. Salek-Ardakani, S. et al. (2003) J. Exp. Med. 198:315.
    11. Jember, A. G. et al. (2001) J. Exp. Med. 193:387.
    12. Demirci, G. et al. (2004) J. Immunol. 172:1691.
    13. Blazar, B.R. et al. (2003) Blood 101:3741.
    14. Higgins, L.M. et al. (1999) J. Immunol. 162:486.
  • Entrez Gene IDs:
    7293 (Human); 22163 (Mouse); 25572 (Rat)
  • Alternate Names:
    ACT-135; ACT35 antigen; ACT35ATC35 antigen; CD134 antigen; CD134; OX40 cell surface antigen; OX40 homologue; OX40L receptor; OX40lymphoid activation antigene ACT35; TAX transcriptionally-activated glycoprotein 1 receptor; tax-transcriptionally activated glycoprotein 1 receptor; TNFRSF4; tumor necrosis factor receptor superfamily member 4; tumor necrosis factor receptor superfamily, member 4; TXGP1LOX40 antigen
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