Recombinant Human OX40/TNFRSF4 Fc Avi-tag Protein, CF

When Recombinant Human OX40 Ligand/TNFSF4 (1054-OX) is immobilized at 0.500 μg/mL (100 μL/well), Biotinylated Recombinant Human OX40/TNFRSF4 Fc Chimera Avi-tag (Catalog # AVI10842) binds with an ED50 of 3.00-15.0 ng/mL.

2 μg/lane of Biotinylated Recombinant Human OX40/TNFRSF4 Fc Chimera Avi-tag Protein (Catalog # AVI10842) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 64-72 kDa and 128-144kDa, respectively.

Product Discontinued

Recombinant Human OX40/TNFRSF4 Fc Avi-tag Protein, CF Summary

• Additional Information: Biotinylated
• Details of Functionality: Measured by its binding ability in a functional ELISA. When Recombinant Human OX40 Ligand/TNFSF4 (Catalog # 1054-OX) is immobilized at 0.500 µg/mL (100 µL/well), Biotinylated Recombinant Human OX40/TNFRSF4 Fc Chimera Avi-tag binds with an ED₅₀ of 3.00-15.0 ng/mL. The biotin to protein ratio is greater than 0.7 as determined by the HABA assay.
• Source: Human embryonic kidney cell, HEK293-derived human OX40/TNFRSF4 protein
  

  Human OX40/TNFRSF4 (Leu29-Ala216) Accession # P43489.1	IEGRMD	Human IgG1 (Pro100-Lys330)	Avi-tag
  N-terminus			C-terminus

• Accession #: P43489.1
• N-terminal Sequence: Leu29
• Structure / Form: Disulfide-linked homodimer, biotinylated via Avi-tag
• Protein/Peptide Type: Recombinant Proteins
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
  • Bioactivity2
• Theoretical MW: 49 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 64-72 kDa, under reducing conditions

Packaging, Storage & Formulations

• 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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 500 μg/mL in PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human OX40/TNFRSF4 Fc Avi-tag Protein, CF

• 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

Background

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). Our Avi-tag Biotinylated human OX40/TNFRSF4 Fc Chimera features biotinylation at a single site contained within the Avi-tag, a unique 15 amino acid peptide. Protein orientation will be uniform when bound to streptavidin-coated surface due to the precise control of biotinylation and the rest of the protein is unchanged so there is no interference in the protein's bioactivity.

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

Bioinformatics

• Uniprot:
  • P43489.1()