Human OX40 Protein, ECD Fc-fusion, Biotinylated, Recombinant

Description

OX40, also known as CD134, ACT35, TXGP1L and TNFRSF4, is a single-pass type I transmembrane glycoprotein of the TNF receptor superfamily (TNFRSF).  OX40 is regarded as a T cell co-stimulatory molecule that coordinates with other membrane­bound co-stimulators including CD28, CD40, CD30, CD27, and 4­1BB/CD137. OX40 is absent on naïve T cells and its expression is induced after engagement of the TCR by antigen presenting cells and co-stimulation by CD40/CD40 ligand and CD28/B7. OX40 was identified as the receptor for OX40 ligand/TNFSF4.  OX40/OX40L-mediated signaling is critical for establishing or reactivating long-term T-cell immunity and breaking immune tolerance in malignancies, leading to a reactivation of anti-tumor immunity. Engagement of OX40 on activated T cells with OX40 ligand on dendritic cells promotes T cell survival and proliferation, prolongs the immune response, and enhances the transition from effector to memory T cells. OX40 also regulates cytokine production from T cells, antigen-presenting cells, natural killer cells, and natural killer T cells. Mutations in the OX40 gene are associated with immunodeficiency 16 (IMD16). Blockade of OX40 signaling with antibody or soluble receptor is efficacious in animal models of allergic airway inflammation, graft­versus­host disease and autoimmune disease. Conversely, stimulating OX40 with an agonistic antibody has shown therapeutic potential for cancer and infectious disease.

  

 

Gene Symbol: OX40; CD134; ACT35; TNFRSF4; IMD16; TXGP1L

 

NCBI Gene ID: 7293

 

Uniprot Entry: P43489

 

Construct Details: The recombinant human OX40-Fc fusion is expressed as a 414 amino acid protein consisting of Leu29 - Ala214 region of OX40 (UniProt accession #P43489)  and a C-terminal Fc from human IgG1, which exists as a dimer/tetramer under non-reducing conditions (see the gel image inserted).  

 

Source: Human cells stably expressing human OX40/CD134 and growing in chemical-defined media with no animal components or antibiotics

 

Deduced Amino Acid Sequence:

LHCVGDTYPSNDRCCHECRPGNGMVSRCSRSQNTVCRPCGPGFYNDVVSSKPCKPCTW

CNLRSGSERKQLCTATQDTVCRCRAGTQPLDSYKPGVDCAPCPPGHFSPGDNQACKPW

TNCTLAGKHTLQPASNSSDAICEDRDPPATQPQETQGPPARPITVQPTEAWPRTSQGP

STRPVEVPGGRASTGTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVD

VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV

SNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEW

ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQK

SLSLSPGK

 

M.W.: Calculated molecular mass (kDa): 45.5; Estimated by SDS-PAGE under reducing condition (kDa): ~65

 

Calculated PI: 8.12

 

Calculated extinction coefficients (M-1 cm-1, at 280nm):  57880

 

Purity: >95% judged by SDS-PAGE under reducing condition (see the gel image above, labeled as "DTT: +")

 

Formulation: Supplied at 0.5 mg/ml in sterile PBSpH7.4 (carrier & preservative free). The purified recombinant protein was labeled with Biotin (3-5 Biotin per molecule) using the standard procedure.

 

Endotoxin Level: <0.1 EU per 1 μg of purified recombinant protein determined by the LAL method

 

Biological Activity: Binds to OX40 ligand and anti-OX40/CD134 monoclonal antibodies (SKU#MAB0103) with high affinity by ELISA (see Technical Data). Blocks OX40 Ligand­induced signaling activity.

 

 

  

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

Restriction: This product is not transferable or re-sellable.  Customer obtain no right to transfer, assign, or sublicense its use rights, or to transfer, resell, package, or otherwise distribute the product, or to use the product for the benefit of any third party or for any commercial purpose.  Customer may only use the product in compliance with applicable local, state and federal laws, regulations and rules.  Customer may not directly or indirectly use the product or allow the transfer, transmission, export or re-export of all or any part of the product in violation of any export control law or regulation of the united states or any other relevant jurisdiction.  Your use of this product constitutes acceptance of the terms of this limited use agreement.  Please refer to our “terms & conditions” for details.  If you are not willing to accept the limitation of this agreement, G&P Biosciences will accept return of the product for a full/partial refund.

 

 

Recombinant OX40-Fc protein (SKU#FCL0103) was immobilized at 50 ng/well on a 96-well NuncMaxisorp plate for ELISA with serially diluted biotinylated anti-OX40/CD134monoclonal antibody, human IgG1 (SKU#MAB0103B). Biotinyalted anti-B7-H3/CD276 (SKU#MAB1755B) human IgG1 was used as a control to demonstrate the specificity of OX40/CD134 binding under the experimental conditions.  The binding curve was fit and apparent K_D was calculated for anti-OX40:OX40 binding to be 132 ± 7 pM.   

 

 

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

 

 

The product is shipped at 4°C. Upon receipt, centrifuge the product briefly before opening the vial. It is recommended to store small aliquots at the temperature below –20°C for long-term storage and the product is stable for 3 months. The undiluted protein can be stored at 4°C for no more than 2 weeks.

 

 

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

 

 

 

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

 

 

1. Eur. J. Immunol. 24:677 (1994).

2. Mol. Cell. Biol. 18:558 (1998)

3. Nat. Med. 7:907 (2001).

4. Immunity 15:445 (2001).

5. J. Exp. Med. 210:1743 (2013).

 

 

 

 

 

Product datasheet (pdf) can be downloaded here: FCL0103B-PDS.pdf

 

 

 

Additional supporting documents, including COA and MSDS are available upon request.

 

 

 

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

Restriction: This product is not transferable or re-sellable.  Customer obtain no right to transfer, assign, or sublicense its use rights, or to transfer, resell, package, or otherwise distribute the product, or to use the product for the benefit of any third party or for any commercial purpose.  Customer may only use the product in compliance with applicable local, state and federal laws, regulations and rules.  Customer may not directly or indirectly use the product or allow the transfer, transmission, export or re-export of all or any part of the product in violation of any export control law or regulation of the united states or any other relevant jurisdiction.  Your use of this product constitutes acceptance of the terms of this limited use agreement.  Please refer to our “terms & conditions” for details.  If you are not willing to accept the limitation of this agreement, G&P Biosciences will accept return of the product for a full/partial refund.