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.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG₁

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human OX40 Ligand/TNFSF4 Antibody

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).

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.

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 deltaSyk 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 “

View 1 more images

Applications for Human OX40 Ligand/TNFSF4 Antibody

Application

Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

2.5 µg/10⁶ cells
Sample: Human mature dentritic cells differentated from human peripheral blood mononuclear cell derived CD14⁺ cells treated with Recombinant Human IL‑4 (Catalog # 204-IL), Recombinant Human GM‑CSF (Catalog # 215-GM), LPS, Recombinant Human TNF‑ alpha (Catalog # 210-TA), and Recombinant Human IL‑1 beta/IL‑1F2 (Catalog # 201-LB)

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed human peripheral blood mononuclear cells stimulated with LPS

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.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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

Godfrey, W.R. et al. (1994) J. Exp. Med. 180:757.
Baum, P.R. et al. (1994) EMBO J. 13:3992.
Al-Shamkhani, A. et al. (1997) J. Biol. Chem. 272:5275.
Kjaergaard, J. et al. (2000) Cancer Res. 60:5514.

Alternate Names

CD252, gp34, TNFSF4

Entrez Gene IDs

7292 (Human); 22164 (Mouse); 89814 (Rat)

Gene Symbol

TNFSF4

UniProt

P23510

Additional OX40 Ligand/TNFSF4 Products

Product Documents for Human OX40 Ligand/TNFSF4 Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human OX40 Ligand/TNFSF4 Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human OX40 Ligand/TNFSF4 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB10541

Key Product Details

Validated by

Species Reactivity

Validated:

Cited:

Applications