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 Mouse OX40 Ligand/TNFSF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1236). The ND50 is typically 0.2-0.6 µg/mL.
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.