Anti-B7-H3/CD276 Antibody, Human IgG1, Biotinylated, Recombinant

Description

B7­H3 (B7 homolog 3), also known as CD276, is a single pass type I transmembrane glycoprotein that belongs to the B7 family of the Ig superfamily. Like other B7 family members, B7-H3 contains one Ig V-like and one Ig C-like domain in the extracellular region. Among the family members, they share about 20-25% amino acid identity. B7-H3 serves as an accessory modulator of T-cell response. It has been reported that B7-H3 can provide both a stimulatory and inhibitory signal to T cells. As a negative regulator, it preferentially affects Th1 responses. B7-H3 may also play an important role in muscle-immune interactions. A longer form of B7-H3 has been identified in human but not in mouse. It is termed 4IgB7­H3 or B7­H3b with two additional Ig­like domains (one V-type and one C-type). B7-H3b appears to be the dominantly expressed form in human and is found on human dendritic cells, activated T, B and NK cells. Human B7­H3b binding to an undefined receptor may inhibit NK cell killing and cytokine release.  It is also involved in late stage osteoblast differentiation. B7-H3 is detected in a variety of cancer types and its expression in some cancers correlates with poor outcome of cancer patients.

 

 

Gene Symbol: B7-H3; CD276; B7H3; B7RP-2; 4Ig-B7-H3; PSEC0249; UNQ309/PRO352

 

NCBI Gene ID: 80381

 

Uniprot Entry: Q5ZPR3

 

Construct Details: Expressed as the combination of a heavy chain (HC) containing VH from anti-B7-H3/CD276 mAb and CH1-3 region of human IgG1 and a light chain (LC) encoding VL from anti-B7-H3 mAb and CL of human kappa light chain.  Exists as a disulfide linked dimer of the HC and LC hetero-dimer under non-reducing condition.

 

Source: Mammalian cells stably expressing anti-B7-H3 heavy chain and light chain (in human IgG1,k format) and growing in chemical-defined media with no animal components. 

 

M.W.: Calculated molecular mass (kDa): 147 (49 kDa for a single HC, 24.5 kDa for a single LC); Estimated by SDS-PAGE under non-reducing condition(kDa): 150

 

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

 

Formulation: Supplied at 1.0 mg/ml in sterile PBS pH7.4 without any carrier protein or preservative (the concentration determined by UV absorption with an A280nm of 1.0 for 0.73 mg/ml of IgG and verified by SDS-PAGE and Coomassie blue staining). The purified recombinant monoclonal antibody was labeled with Biotin (3-5 Biotin per IgG) 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 human B7-H3 and B7-H3b proteins (SKU#FCL1198, FCL1662) with high affinity (K_D < 1 nM) by ELISA (with the same specificity as anti-B7-H3 rabbit IgG, SKU#MAB1753 - see Technical Data). Recognizes cell surface human B7-H3/CD276 by flow cytometry, IHC staining and fluorescent microscopy. Enhances T cell-dependent anti-tumor immunity.

 

 

  

 

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 B7-H3-Fc protein (SKU#FCL1198) was immobilized at 50 ng/well on a 96-well NuncMaxisorp plate for ELISA with serially diluted biotinylated anti-B7-H3/CD276 monoclonal antibody, human IgG1 (SKU#MAB1755B). Biotinylated anti-OX40 (SKU#MAB0103B) human IgG1 was used as a control to demonstrate the specificity of B7-H3/CD276 binding under the experimental conditions.  The binding curve was fit and apparent K_D was calculated for anti-B7-H3:B7-H3 binding to be 163 ± 7 pM.   

 

Recombinant B7-H3b-Fc protein (SKU#FCL1662) was immobilized at 50 ng/well on a 96-well NuncMaxisorp plate for ELISA with serially diluted biotinylated anti-B7-H3/CD276 monoclonal antibody, human IgG1 (SKU#MAB1755B). Biotinylated anti-OX40 (SKU#MAB0103B) human IgG1 was used as a control to demonstrate the specificity of B7-H3b binding under the experimental conditions.  The binding curve was fit and apparent K_D was calculated for anti-B7-H3:B7-H3b binding to be 152 ± 8 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. Nat. Immunol. 2:269 (2001). 

2. Nat. Immunol. 2:203 (2001).  

3. J. Immunol. 168:6294 (2002).  

4. J. Immunol. 172:2352 (2004).  

5. Proc. Natl. Acad. Sci. 101:12969 (2004).

 

 

 

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

 

 

Product datasheet (pdf) can be downloaded here: MAB1755B-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.