Mouse OX40 Ligand/TNFSF4 Antibody

(1 citations)   
  • Species Reactivity
    Mouse
  • Specificity
    Detects mouse OX40 Ligand/TNFSF4 in direct ELISAs and Western blots. In Western blots, approximately 5% cross-reactivity with recombinant human (rh) OX40 Ligand, rhLIGHT, recombinant mouse (rm) Fas Ligand and less than 1% cross-reactivity with rmTNF‑ alpha, rhGITR Ligand, rhTRANCE, and rhVEGI is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant mouse OX40 Ligand/TNFSF4
    Gln49-Leu198
    Accession # P43488
  • 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
  • Western Blot
    0.1 µg/mL
    Recombinant Mouse OX40 Ligand/TNFSF4 (Catalog # 1236-OX)
  • Immunocytochemistry
    5-15 µg/mL
    See below
  • 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.
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
Immunocytochemistry
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 CD3 epsilon 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 Mouse OX40 Ligand/TNFSF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1236). The ND50 is typically 0.2-0.6 µg/mL.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 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 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).

  • References:
    1. Baum, P.R. et al. (1994) EMBO J. 13:3992.
    2. Gramaglia, I. et al. (1999) J. Immunol. 161:6510.
    3. Stuber, E. et al. (1995) Immunity 2:507.
    4. Malstrom, V. et al. (2001) J. Immunol. 166:6972.
    5. Maxwell, J.R. et al. (2000) J. Immunol. 164:107.
    6. Weinberg, A.D. (2002) Trends Immunol. 23:102.
  • 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.

1 Citations: Showing 1 - 1

  1. Group 2 Innate Lymphoid Cells Promote an Early Antibody Response to a Respiratory Antigen in Mice
    J Immunol, 2016;0(0):.
    Species: Mouse
    Sample Type: Whole Cells
    Application: Neut
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