Human OX40 Ligand/TNFSF4 Antibody

Catalog # Availability Size / Price Qty
MAB10541-SP
MAB10541-100
MAB10541-500
Detection of OX40 Ligand/ TNFSF4 in Human Mature Dendritic Cells by Flow Cytometry.
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Product Details
Citations (8)
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Human OX40 Ligand/TNFSF4 Antibody Summary

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 either lyophilized or 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.

Scientific Data

Flow Cytometry Detection of OX40 Ligand/ TNFSF4 antibody in Human Mature Dendritic Cells antibody by Flow Cytometry. View Larger

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-a (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 antibody in Human PBMCs by Immunocytochemistry (ICC). View Larger

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.

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

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.

Flow Cytometry Detection of Mouse OX40 Ligand/TNFSF4 by Flow Cytometry View Larger

Detection of Mouse OX40 Ligand/TNFSF4 by Flow Cytometry Th2 polarization induced by SEA is mediated via Dectin-2 and Syk signaling in vivo.(A–D) WT or CD11c delta Syk mice were injected with SEA (panel A, B), SEA delta alpha -1/ omega -1 (panel C), or omega -1 (panel D) in the hind footpad, and draining pLNs were analyzed 7 d later. (A, C, D) pLN cells were restimulated with PMA/Ionomycin in the presence of brefeldin A, and CD4+ T cells were stained for indicated intracellular cytokines and percentage cytokine-positive CD4+ T cells enumerated. Based on these data, the ratio between the percentage IL-4+ over IFN-gamma + CD4+ T cells was determined as a measure for overall skewing towards Th2. (B) pLN cells were restimulated with SEA for 72 h, and cytokine levels in culture supernatants were determined. (A–D) Bar graphs represent mean ± SEM of 5 to 6 mice per group and are representative of 2 (panel A–C) or 1 (panel D) experiment. (E) BMDCs cultured from BM from WT, Dectin-1−/−, or Dectin-2−/− mice were pulsed overnight with SEA delta alpha -1/ omega -1 and injected into hind footpads after which CD4+ T-cell responses were analyzed as in panel A. Representative flow cytometry plots of intracellular staining of CD4+ T cells are depicted, of which the data are enumerated in bar graphs representing mean ± SEM of 2 independent experiments with 3 to 6 mice per group. (F) SEA binding and uptake was determined as in Fig 4F. (G) ROS production by indicated BMDCs was determined as described in Fig 5E, 1 h after stimulation with SEA delta alpha -1/ omega -1. Based on MFI, bar graphs represent fold change relative to control condition, which is set to 1. (H) BMDCs were stimulated as indicated for 18 h after which expression of OX40L was analyzed by flow cytometry. Representative plots are depicted, of which the data are enumerated in bar graphs and shown as fold change relative to control condition, which is set to 1, based on percentage positive cells. (F–H) Bar graphs represent mean of duplicates or triplicates ± SEM of 2 independent experiments. “*”: P < 0.05; “**” and “##”: P < 0.01; “***” and “###”: P < 0.001 for significant differences with the control conditions (*) or between-test condition (#) based on unpaired analysis (unpaired Student t test). Underlying data can be found in S1 Data. omega -1, omega-1; BMDC, bone marrow–derived DC; CD4, cluster of differentiation 4; H2-DCFDA, 2',7'-dichlorodihydrofluorescein diacetate; IFN-gamma, interferon gamma ; IL-4, interleukin 4; MFI, mean fluorescence intensity; nd/pLN, nondraining/popliteal lymph node; OX40L, OX40 ligand; PBS, phosphate buffered saline; PMA, phorbol 12-myristate 13-acetate; ROS, reactive oxygen species; SEA, soluble egg antigen; Syk, spleen tyrosine kinase; Th2, T helper 2; WT, wild-type. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pbio.2005504), licensed under a CC-BY license. Not internally tested by R&D Systems.

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
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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

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Citations for Human OX40 Ligand/TNFSF4 Antibody

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.

8 Citations: Showing 1 - 8
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  1. Predictive biomarkers for the responsiveness of recurrent glioblastomas to activated killer cell immunotherapy
    Authors: S Hwang, J Lim, H Kang, JY Jeong, JG Joung, J Heo, D Jung, K Cho, HJ An
    Cell & bioscience, 2023-01-24;13(1):17.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  2. Priming Leukemia with 5-Azacytidine Enhances CAR T Cell Therapy
    Authors: Ning Xu, Benjamin Tse, Lu Yang, Tiffany C Y Tang, Michelle Haber, Kenneth Micklethwaite et al.
    ImmunoTargets and Therapy
  3. OX40L/OX40 axis impairs follicular and natural Treg function in human SLE
    Authors: C Jacquemin, JF Augusto, M Scherlinge, N Gensous, E Forcade, I Douchet, E Levionnois, C Richez, E Lazaro, P Duffau, ME Truchetet, J Seneschal, L Couzi, JL Pellegrin, JF Viallard, T Schaeverbe, V Pascual, C Contin-Bor, P Blanco
    JCI Insight, 2018-12-20;3(24):.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  4. Dectin-1/2-induced autocrine PGE2 signaling licenses dendritic cells to prime Th2 responses
    Authors: MMM Kaisar, M Ritter, C Del Fresno, HS Jónasdótti, AJ van der Ha, LR Pelgrom, G Schramm, LE Layland, D Sancho, C Prazeres d, M Giera, M Yazdanbakh, B Everts
    PLoS Biol., 2018-04-18;16(4):e2005504.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  5. Soluble OX40L is associated with presence of autoantibodies in early rheumatoid arthritis
    Authors: Julie K Laustsen, Tue K Rasmussen, Kristian Stengaard-Pedersen, Kim Hørslev-Petersen, Merete L Hetland, Mikkel Østergaard et al.
    Arthritis Research & Therapy
  6. 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, 2012-06-29;7(6):e39576.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  7. B7-CD28 costimulatory signals control the survival and proliferation of murine and human gammadelta T cells via IL-2 production.
    J. Immunol., 2012-06-25;189(3):1202-8.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  8. 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, 2004-09-16;105(2):750-8.
    Species: Human
    Sample Types: Cell Culture Supernates
    Applications: Neutralization

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