HEK293 human embryonic kidney cell line transfected with either (A) human OX40/TNFRSF4 or (B) irrelevant protein, and eGFP, was stained with Rabbit anti-human OX40/TNFRSF4 monoclonal antibody (Catalog # MAB10563) followed by Allophycocyanin-conjugated anti-Rabbit IgG Secondary Antibody (F0111). Quadrant markers were set based on control antibody staining (MAB1050, data not shown). Staining was performed using our Staining Membrane-Associated Proteins protocol.

CXCL8/IL-8 Secretion Induced by OX40L/TNFSF4 and Neutralization by Human OX40/TNFRSF4 Antibody.

Recombinant Human OX40L/TNFSF4 (1054-OX) stimulates CXCL8/IL-8 secretion in the HT1080 human fibrosarcoma cell line transfected with human OX40/TNFRSF4 in a dose-dependent manner (orange line), as measured by the Human CXCL8/IL-8 Quantikine ELISA Kit (D8000C). CXCL8/Il-8 Secretion elicited by Recombinant Human OX40L/TNFSF4 (20 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human OX40/TNFRSF4 Monoclonal Antibody (Catalog # MAB10563). The ND₅₀ is typically 25-250 ng/mL.

Applications for Human OX40/TNFRSF4 Antibody

Application

Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/10⁶ cells
Sample: HEK293 Human Cell Line transfected with Human OX40/TNFRSF4 and eGFP

Neutralization

Measured by its ability to neutralize OX40L/TNFSF4-induced IL-8 secretion in the HT1080 human fibrosarcoma cell line transfected with human OX40/TNFRSF4. The Neutralization Dose (ND₅₀) is typically 25-250 ng/mL in the presence of 20 ng/mL Recombinant Human OX40L/TNFSF4.

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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

OX40 (CD134; TNFRSF4) is a T cell co-stimulatory molecule of the TNF receptor superfamily that coordinates with other co-stimulators (CD28, CD40, CD30, CD27 and 4-1BB) to manage the activation of the immune response (1-3). Human OX40 is a 48 kDa type I transmembrane glycoprotein with a 28 amino acid (aa) signal sequence, a 185 aa extracellular domain (ECD) that contains a cysteine-rich region, a 20 aa transmembrane segment, and a 41 aa cytoplasmic domain (4). The ECD of human OX40 shares 63% sequence identity with the ECD of mouse and rat OX40. OX40 is up-regulated on CD4⁺ and CD8⁺ T cells upon engagement of the TCR by antigen presenting cells along with co-stimulation by CD40-CD40 Ligand and CD28-B7 (5, 6). OX40 Ligand is primarily expressed on antigen presenting cells (5). OX40 Ligand engagement of OX40 on activated CD4⁺ T cells results in increased T cell survival, proliferation, and cytokine production. It also inhibits the conversion of effector T cells into immunosuppressive regulatory T cells (Tregs) and can promote the maintenance of and recall response in memory T cells (3, 7-10). OX40 is constitutively expressed on Tregs and enhances the sensitivity of Tregs to IL-2, thus promoting Treg proliferation. OX40 has also been shown to decrease the cells’ immunosuppressive activity on effector T cells (11-14). OX40-OX40 Ligand signaling is involved in allergic airway inflammation, graft-versus-host disease and autoimmune disease (6, 15, 16). Mutations in OX40 and OX40 Ligand are associated with cardiovascular disease (17, 18).

References

Hori, T. (2006) Int. J. Hematol. 83:17.
Latza, U. et al. (1994) Eur. J. Immunol. 24:677.
Salek-Ardakani, S. et al. (2003) J. Exp. Med. 198:315.
al-Shamkhani, A. et al. (1996) Eur. J. Immunol. 26:1695.
Moran, A.E. et al. (2013) Curr. Opin. Immunol. 25:230.
Gramaglia, I. et al. (1998) J. Immunol. 161:6510.
Xiao, X. et al. (2008) J. Immunol. 181:3193.
So, T. and M. Croft (2007) J. Immunol. 179:1427.
Mousavi, S.F. et al. (2008) J. Immunol. 181:5990.
Bansal-Pakala, P. et al. (2001) Nat. Med. 7:907.
Piconese, S. et al. (2010) Eur. J. Immunol. 40:2902.
Griseri, T. et al. (2010) J. Exp. Med. 207:699.
Xiao, X. et al. (2012) J. Immunol. 188:892.
Vu, M.D. et al. (2007) Blood 110:2501.
Damayanti, T. et al. (2010) Am. J. Respir. Crit. Care Med. 181:688.
Xiao, X. et al. (2012) Nat. Immunol. 13:981.
Nakano, M. et al. (2010) Cardiovasc. Res. 88:539.
Ishii, N. et al. (2010) Adv. Immunol. 105:63.
Godfrey, W.R. et al. (1994) J. Exp. Med. 180:757

Alternate Names

ACT-135, CD134, Ly-70, TNFRSF4, Txgp1, TXGP1L

Entrez Gene IDs

7293 (Human); 22163 (Mouse); 25572 (Rat); 102145978 (Cynomolgus Monkey)

Gene Symbol

TNFRSF4

UniProt

P34389

Additional OX40 Products

Product Documents for Human OX40/TNFRSF4 Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

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Product Specific Notices for Human OX40/TNFRSF4 Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human OX40/TNFRSF4 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB10563

Key Product Details

Species Reactivity

Applications

Label

Antibody Source

View all OX40 Antibodies »

Product Specifications

Immunogen

Specificity

Clonality

Host

Isotype

Endotoxin Level

Scientific Data Images for Human OX40/TNFRSF4 Antibody

Detection of OX40/TNFRSF4 in HEK293 Human Cell Line transfected with Human OX40/TNFRSF4 and eGFP by Flow Cytometry