Human OX40/TNFRSF4 Antibody

  
  • Specificity
    Detects human OX40/TNFRSF4 in direct ELISAs and Western blots. In Western blots, no cross-reactivity with recombinant human (rh) 4-1BB, rhBAFF R, rhCD27, rhCD30, rhCD40, rhDR3, rhDR6, rhEDAR, rhFAS, rhGITR, rhHVEM, rhLT beta R, rhNGF R, rhRANK, rhRELT, rhTAJ, rhTNF RI, rhTNF RII, rhTRAIL, rhTRAIL R4, rhTRAIL R3, rhTWEAK R, or rhXEDAR is observed.
  • Source
    Monoclonal Rat IgG2A Clone # 443318
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human OX40/TNFRSF4
    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
    1 µg/mL
    Recombinant Human OX40/TNFRSF4 Fc Chimera (Catalog # 3388-OX)
  • 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 OX40/TNFRSF4 in Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells (PBMCs) were stained with Mouse Anti-Human CD3 epsilon APC‑conjugated Monoclonal Antibody (Catalog # FAB100A) and either (A) Rat Anti-Human OX40/TNFRSF4 Monoclonal Antibody (Catalog # MAB3388) or (B) Rat IgG2A Isotype Control (Catalog # MAB006) followed by Phycoerythrin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0105B). View our protocol for Staining Membrane-associated Proteins
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: OX40/TNFRSF4

OX40 (CD134; TNFRSF4) is a T cell co‑stimulatory molecule of the TNF receptor superfamily that coordinates with other membrane-bound co‑stimulators 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 co‑stimulation 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; CD134; TNFRSF4; ACT35 antigen; ACT35ATC35 antigen; CD134; CD134 antigen; OX40 cell surface antigen; OX40 homologue; OX40L receptor; OX40lymphoid activation antigene ACT35; TAX transcriptionally-activated glycoprotein 1 receptor; tax-transcriptionally activated glycoprotein 1 receptor; tumor necrosis factor receptor superfamily member 4; tumor necrosis factor receptor superfamily, member 4; TXGP1LOX40 antigen
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