Human OX40/TNFRSF4 Antibody

Catalog # Availability Size / Price Qty
MAB10543-100
MAB10543-SP
Detection of OX40/TNFRSF4 in HEK293 Human Cell Line Transfected with Human OX40/TNFRSF4 and eGFP by Flow Cytometry.
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Human OX40/TNFRSF4 Antibody Summary

Species Reactivity
Human
Specificity
Detects human OX40/TNFRSF4 in direct ELISAs
Source
Monoclonal Mouse IgG2B Clone # 977974
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Mouse myeloma cell line NS0-derived recombinant human OX40/TNFRSF4
Leu29-Ala216
Accession # P43489
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
Flow Cytometry
0.25 µg/106 cells
See below
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 
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 0.6-3.0 µg/mL in the presence of 10 ng/mL Recombinant Human OX40L/TNFSF4.

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

Flow Cytometry Detection of OX40/TNFRSF4 in HEK293 Human Cell Line Transfected with Human OX40/TNFRSF4 and eGFP by Flow Cytometry. View Larger

Detection of OX40/TNFRSF4 in HEK293 Human Cell Line Transfected with Human OX40/TNFRSF4 and eGFP by Flow Cytometry. HEK293 human embryonic kidney cell line transfected with (A) human OX40/TNFRSF4 or (B) irrelevant transfectants and eGFP was stained with Mouse Anti-Human OX40/TNFRSF4 Monoclonal Antibody (Catalog # MAB10543) followed by APC-conjugated Anti-Mouse IgG Secondary Antibody(Catalog # F0101B). Quadrant markers were set based on control antibody staining (Catalog # MAB0041). View our protocol for Staining Membrane-associated Proteins.

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

CXCL8/Il-8 Secretion Induced by OX40L/TNFSF4 and Neutralization by Human OX40/TNFRSF4 Antibody. Recombinant Human OX40L/TNFSF4 (Catalog # 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 (Catalog # D8000C). CXCL8/Il-8 Secretion elicited by Recombinant Human OX40L/TNFSF4 (10 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human OX40/TNFRSF4 Monoclonal Antibody (Catalog # MAB10543). The ND50 is typically 0.6-3.0 µg/mL.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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/TNFRSF4

OX40 (CD134; TNFRSF4) is a T cell co‑stimulatory molecule of the TNF receptor superfamily that coordinates with other membrane-bound co‑stimulators such as CD28, CD40, CD30, CD27 and 4-1BB (1‑3). OX40 is expressed on naïve CD4+ T cells only after engagement of the TCR by antigen presenting cells (APC; dendritic and B cells), and co‑stimulation by CD40/CD40 ligand and CD28/B7. It is maximal at 2‑5 days post activation, or 4 hours post reactivation of memory T cells (3‑6). 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 has four TNFR-Cys repeats and an O-glycosylated hinge region, a 20 aa transmembrane segment, and a 41 aa cytoplasmic domain (3). The ECD of human OX40 shows 71%, 68%, 67%, 64% and 64% aa identity with feline, canine, rabbit, mouse and rat OX40 ECD, respectively. Engagement of OX40 on activated CD4+ T cells by OX40 ligand on activated dendritic cells promotes T cell survival and proliferation, prolongs the immune response, and enhances the number of cells making the transition from effector to memory T cells (1‑6). OX40 signal transduction includes binding TNF receptor-associated factors (TRAFs), and activating NF kappa B and PI3 kinase to enhance expression of cytokines, antiapoptotic Bcl-2 family members, survivin and the chemokine receptor CXCR5 (5‑8). CXCR5 promotes T cell migration to germinal centers to deliver B cell help (5). Studies using knockout or transgenic mice, and agonistic or blocking antibodies, show that OX40/OX40L interaction is critical for establishing or reactivating memory T cells and breaking immune tolerance (9). Blockade of OX40 engagement is efficacious in animal models of allergic airway inflammation, graft-versus-host disease and autoimmune disease (10‑14).

 

References
  1. Salek-Ardakani, S. and M. Croft (2006) Vaccine 24:872.
  2. Hori, T. (2006) Int. J. Hematol. 83:17.
  3. Latza, U. et al. (1994) Eur. J. Immunol. 24:677.
  4. Murata, K. et al. (2000) J. Exp. Med. 191:365.
  5. Fillatreau, S and D. Gray (2003) J. Exp. Med. 197:195.
  6. Gramaglia, I. et al. (1998) J. Immunol. 161:6510.
  7. Rogers, P.R. et al. (2001) Immunity 15:445.
  8. Song, J. et al. (2005) Immunity 22:621.
  9. Bansal-Pakala, P. et al. (2001) Nat. Med. 7:907.
  10. Salek-Ardakani, S. et al. (2003) J. Exp. Med. 198:315.
  11. Jember, A. G. et al. (2001) J. Exp. Med. 193:387.
  12. Demirci, G. et al. (2004) J. Immunol. 172:1691.
  13. Blazar, B.R. et al. (2003) Blood 101:3741.
  14. Higgins, L.M. et al. (1999) J. Immunol. 162:486.
Entrez Gene IDs
7293 (Human); 22163 (Mouse); 25572 (Rat); 102145978 (Cynomolgus Monkey)
Alternate Names
ACT-135; ACT35 antigen; ACT35ATC35 antigen; CD134 antigen; CD134; Ly-70; OX40 cell surface antigen; OX40 homologue; OX40; OX40L receptor; OX40lymphoid activation antigene ACT35; TAX transcriptionally-activated glycoprotein 1 receptor; tax-transcriptionally activated glycoprotein 1 receptor; TNFRSF4; tumor necrosis factor receptor superfamily member 4; tumor necrosis factor receptor superfamily, member 4; Txgp1; TXGP1L; TXGP1LOX40 antigen

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Isotype Controls

Reconstitution Buffers

Secondary Antibodies

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