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Mouse OX40 Ligand/TNFSF4 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF1236

R&D Systems, part of Bio-Techne
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AF1236
AF1236-SP

Key Product Details

Species Reactivity

Validated:

Mouse

Cited:

Mouse

Applications

Validated:

Immunocytochemistry, Neutralization, Western Blot

Cited:

Neutralization

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse OX40 Ligand/TNFSF4
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 antibody in Mouse Splenocytes by Immunocytochemistry (ICC).

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.

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

Western Blot

0.1 µg/mL
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.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

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.

Reconstitution Buffer Available:
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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.

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.

Alternate Names

CD252, gp34, TNFSF4

Entrez Gene IDs

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

Gene Symbol

TNFSF4

UniProt

Additional OX40 Ligand/TNFSF4 Products

Product Documents for Mouse OX40 Ligand/TNFSF4 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Mouse OX40 Ligand/TNFSF4 Antibody

For research use only

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