Detects mouse OX40 Ligand/TNFSF4 in direct ELISAs and Western blots. In direct ELISAs and Western blots, no cross-reactivity with recombinant human (rh) APRIL, rhGITR Ligand, rhLIGHT, rhLT alpha 1/ beta 2, rhLT alpha 2/ beta 1, recombinant cotton rat TNF‑ alpha, recombinant mouse (rm) TNF‑ alpha, rmTRAIL, rhTRANCE, rmTRANCE, rmTWEAK, or rhVEGI is observed.
Monoclonal Rat IgG2A Clone # 182601
Protein A or G purified from hybridoma culture supernatant
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.
<0.10 EU per 1 μg of the antibody by the LAL method.
Recombinant Mouse OX40 Ligand/TNFSF4 (Catalog # 1236-OX) under non-reducing conditions only
Measured by its ability to neutralize OX40 Ligand/TNFSF4-induced IL‑2 secretion in mouse T cells. The Neutralization Dose (ND50) is typically 0.01-0.05 µg/mL in the presence of 30 ng/mL Recombinant Mouse OX40 Ligand/TNFSF4, 10 µg/mL of 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.
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 (30 ng/mL) is neutralized (green line) by increasing concentrations of Rat Anti-Mouse OX40 Ligand/TNFSF4 Monoclonal Antibody (Catalog # MAB1236). The ND50 is typically 0.01-0.05 µg/mL.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
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) 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 mouse OX40L share 46% sequence identity at the amino acid level (1). 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) which 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 upregulated 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).
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