Human OX40 Ligand/TNFSF4 Antibody MAB10541-100: R&D Systems

Human OX40 Ligand/TNFSF4 Antibody

(3 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human OX40 Ligand in direct ELISAs. In direct ELISAs, no cross-reactivity with recombinant human (rh) APRIL, rhBAFF, rhCD27 Ligand, recombinant mouse (rm) CD27 Ligand, rhCD30 Ligand, rmCD30 Ligand, rhCD40 Ligand, rmEDA, rhFas Ligand, rmFas Ligand, rhGITR Ligand, recombinant cotton rat TNF-alpha, rhTNF-alpha, rmTNF-alpha, recombinant porcine TNF-alpha, recombinant rat TNF-alpha, rhTRAIL, rhTRANCE, rmTRANCE, or rhVEGF is observed.
  • Source
    Monoclonal Mouse IgG1 Clone # 159403
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human OX40 Ligand
    Gln51-Leu183
    Accession # P23510
  • 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
  • Flow Cytometry
    2.5 µg/106 cells
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    8-25 µg/mL
    See below
  • Neutralization
    Measured by its ability to neutralize OX40 Ligand/TNFSF4-induced IL-8 secretion in the HT1080 human fibrosarcoma cell line transfected with human OX40. Muller, N. et al. (2008) FEBS Journal 275:2296. The Neutralization Dose (ND50) is typically 0.1-0.6 μg/mL in the presence of 10 ng/mL Recombinant Human OX40 Ligand/TNFSF4.
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 Ligand/
TNFSF4 in Human Mature Dendritic Cells by Flow Cytometry.

Human mature dentritic cells differentated from human peripheral blood mononuclear cell derived CD14+ cells treated with 20 ng/mL Recombinant Human IL‑4 (Catalog # 204-IL) and 50 ng/mL Recombinant Human GM‑CSF (Catalog # 215-GM) for 7 days and 1 μg/mL LPS, 20 ng/mL Recombinant Human TNF‑ alpha (Catalog # 210-TA), and 20 ng/mL Recombinant Human IL‑1 beta /IL‑1F2 (Catalog # 201-LB) for last 24 hours were stained with Mouse Anti-Human OX40 Ligand/TNFSF4 Monoclonal Antibody(Catalog # MAB10541, filled histogram) or isotype control (Catalog # MAB002, open histogram), followed by Goat F(ab)2 Anti-Mouse IgG (H+L) Allophycocyanin (Catalog # F0101B).

Immunocytochemistry
OX40 Ligand/TNFSF4 in Human PBMCs.

OX40 Ligand/TNFSF4 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) stimulated with LPS using Mouse Anti-Human OX40 Ligand/
TNFSF4 Monoclonal Antibody (Catalog # MAB10541) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (orange; Catalog # NL007) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

IL‑8 Secretion Induced by OX40 Ligand/TNFSF4 and Neutralization by Human OX40 Ligand/TNFSF4 Antibody.

Recombinant Human OX40 Ligand/TNFSF4 (Catalog # 1054-OX) induces IL‑8 secretion in the HT1080 human fibrosarcoma cell line transfected with human OX40 in a dose-dependent manner (orange line) as measured by Human IL‑8 Duoset (Catalog # DY208). IL‑8 Secretion elicited by Recombinant Human OX40 Ligand/TNFSF4 (10 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human OX40 Ligand/TNFSF4 Monoclonal Antibody (Catalog # MAB10541). The ND50 is typically 0.1-0.6  μg/mL.

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 Ligand/TNFSF4

OX40 Ligand (OX40L), also known as gp34, is a type II transmembrane glycoprotein designated TNFSF4 within the TNF superfamily. Human OX40L cDNA encodes a 183 amino acids (aa) polypeptide with an amino-terminal cytoplasmic domain (aa 1-23) and a carboxy-terminal extracellular domain (aa 51-183). It shares 46% aa sequence identity with the mouse counterpart. OX40L is expressed on the surface of activated B cells, T cells, dendritic cells and endothelial cells. Like other TNF superfamily members, membrane-bound OX40 Ligand exists as a homotrimer. OX40L binds to OX40 (CD134), a member of the TNF receptor superfamily that is expressed predominantly on activated CD4+ T cells. OX40 Ligand is one of the group of co-stimulatory molecules in the immune system that includes B7, CD40 Ligand, CD30 Ligand, CD27 Ligand and 4-1BB Ligand. OX40 appears as a late activation-induced T cell surface antigen, and its major function of OX40-OX40L interaction may be to transmit a late co-stimulatory signal to promote the survival and proliferation of activated CD4+ T cells and prolong the immune response. Engagement of OX40 on activated T cells in situ in tumors has been shown to augment immune responses and subsequent tumor regression.

  • References:
    1. Godfrey, W.R. et al. (1994) J. Exp. Med. 180:757.
    2. Baum, P.R. et al. (1994) EMBO J. 13:3992.
    3. Al-Shamkhani, A. et al. (1997) J. Biol. Chem. 272:5275.
    4. Kjaergaard, J. et al. (2000) Cancer Res. 60:5514.
  • Entrez Gene IDs:
    7292 (Human); 22164 (Mouse); 89814 (Rat)
  • Alternate Names:
    CD134 ligand; CD134L; CD252 antigen; CD252; Glycoprotein Gp34; gp34; OX40 antigen ligand; OX40 Ligand; OX40L; OX-40L; OX4OL; TAX transcriptionally-activated glycoprotein 1; tax-transcriptionally activated glycoprotein 1 (34kD); tax-transcriptionally activated glycoprotein 1, 34kD; TNFSF4; tumor necrosis factor (ligand) superfamily, member 4; tumor necrosis factor ligand superfamily member 4; TXGP1
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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Species
Applications
Sample Type
  1. Activated Human CD4+CD45RO+ Memory T-Cells Indirectly Inhibit NLRP3 Inflammasome Activation through Downregulation of P2X7R Signalling.
    Authors: Beynon V, Quintana FJ, Weiner HL
    PLoS ONE, 2012;7(6):e39576.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  2. B7-CD28 costimulatory signals control the survival and proliferation of murine and human gammadelta T cells via IL-2 production.
    J. Immunol., 2012;189(3):1202-8.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  3. Cord blood CD4(+)CD25(+)-derived T regulatory cell lines express FoxP3 protein and manifest potent suppressor function.
    Authors: Godfrey WR, Spoden DJ, Ge YG, Baker SR, Liu B, Levine BL, June CH, Blazar BR, Porter SB
    Blood, 2005;105(2):750-8.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: Neut
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