OX40 Ligand (OX40L), also known as gp34, is a type II transmembrane glycoprotein belonging to the TNF superfamily. Murine OX40L cDNA encodes a 198 amino acid (aa) residue protein comprised of a 28 aa N-terminal cytoplasmic domain, a 20 aa transmembrane segment, and a 150 aa C-terminal extracellular domain (1). Human and murine OX40L share 46% sequence identity at the amino acid level (1). The OX40L is expressed on activated antigen presenting cells such as B cells, macrophages, dendritic cells, and on endothelial cells at the site of inflammation. The receptor for OX40L is OX40 (CD134) that is expressed predominantly on activated CD4+ T cells. Expression of OX40 is transient following engagement of T cell receptors (2). Ligation of OX40L by OX40 stimulates proliferation and differentiation of activated B cells, and increases immunoglobulin secretion (3, 4). The expression of OX40L on B cells is up-regulated by CD40 ligation (3). Engagement of the OX40-OX40L system has co-stimulatory effects on T cells by stimulating the production of cytokines by T helper cells and increasing the survival of memory T cells (2, 5). Blocking of the OX40-OX40L interaction in vitro inhibits co-stimulation resulting in decreased T cell proliferation and adhesion of T cells to endothelial cells. Inhibition of the OX40-OX40L interaction in disease models has beneficial effects in acute graft-versus-host disease, inflammatory bowel disease, and decreases the development of collagen-induced arthritis and experimental leishmaniasis (6).
Mouse OX40 Ligand/TNFSF4 Antibody
R&D Systems | Catalog # AF1236
Key Product Details
Species Reactivity
Validated:
Mouse
Cited:
Mouse
Applications
Validated:
Western Blot, Neutralization, Immunocytochemistry
Cited:
Neutralization
Label
Unconjugated
Antibody Source
Polyclonal Goat IgG
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Product Specifications
Immunogen
Mouse myeloma cell line NS0-derived recombinant mouse OX40 Ligand/TNFSF4
Gln49-Leu198
Accession # P43488
Gln49-Leu198
Accession # P43488
Specificity
Detects mouse OX40 Ligand/TNFSF4 in direct ELISAs and Western blots. In direct ELISAs, approximately 5% cross-reactivity with recombinant human OX40 Ligand is observed.
Clonality
Polyclonal
Host
Goat
Isotype
IgG
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Scientific Data Images for Mouse OX40 Ligand/TNFSF4 Antibody
OX40 Ligand/TNFSF4 in Mouse Splenocytes.
OX40 Ligand/TNFSF4 was detected in immersion fixed mouse splenocytes using Goat Anti-Mouse OX40 Ligand/TNFSF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1236) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to the plasma membrane and cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.IL‑2 Secretion Induced by OX40 Ligand/TNFSF4 and Neutralization by Mouse OX40 Ligand/TNFSF4 Antibody.
In the presence of a cross-linking antibody, Mouse polyHistidine Monoclonal Antibody (10 µg/mL, Catalog # MAB050) and sub-optimal amounts of Mouse CD3e Monoclonal Antibody (Catalog # MAB484), Recombinant Mouse OX40 Ligand/TNFSF4 (Catalog # 1236-OX) stimulates IL-2 secretion in mouse T cells in a dose-dependent manner (orange line), as measured by the Mouse IL-2 Quantikine ELISA Kit (Catalog # M2000). Under these conditions, IL-2 secretion elicited by Recombinant Mouse OX40 Ligand/TNFSF4 (100 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse OX40 Ligand/TNFSF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1236). The ND50 is typically 0.2-0.6 µg/mL.Applications for Mouse OX40 Ligand/TNFSF4 Antibody
Application
Recommended Usage
Immunocytochemistry
5-15 µg/mL
Sample: Immersion fixed mouse splenocytes
Sample: Immersion fixed mouse splenocytes
Western Blot
0.1 µg/mL
Sample: Recombinant Mouse OX40 Ligand/TNFSF4 (Catalog # 1236-OX)
Sample: Recombinant Mouse OX40 Ligand/TNFSF4 (Catalog # 1236-OX)
Neutralization
Measured by its ability to neutralize OX40 Ligand/TNFSF4-induced IL‑2 secretion in mouse T cells. The Neutralization Dose (ND50) is typically 0.2-0.6 µg/mL in the presence of 100 ng/mL Recombinant Mouse OX40 Ligand/TNFSF4, 10 µg/mL a cross-linking antibody, Mouse polyHistidine Monoclonal Antibody, and sub-optimal amounts of Mouse CD3 epsilon Monoclonal Antibody.
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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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.
Calculators
Background: OX40 Ligand/TNFSF4
References
- Baum, P.R. et al. (1994) EMBO J. 13:3992.
- Gramaglia, I. et al. (1999) J. Immunol. 161:6510.
- Stuber, E. et al. (1995) Immunity 2:507.
- Malstrom, V. et al. (2001) J. Immunol. 166:6972.
- Maxwell, J.R. et al. (2000) J. Immunol. 164:107.
- Weinberg, A.D. (2002) Trends Immunol. 23:102.
Alternate Names
CD252, gp34, TNFSF4
Gene Symbol
TNFSF4
UniProt
Additional OX40 Ligand/TNFSF4 Products
Product Documents for Mouse OX40 Ligand/TNFSF4 Antibody
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Mouse OX40 Ligand/TNFSF4 Antibody
For research use only
Citations for Mouse OX40 Ligand/TNFSF4 Antibody
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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