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.

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.

 

Product Details

 

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

 

NCBI Gene ID: 80381

 

Uniprot Entry: Q5ZPR3

 

Construct Details: The recombinant human B7-H3b-Fc fusion is expressed as a 666 amino acid protein consisting of Leu29 - Ala466 region of B7-H3b or 4IgB7-H3 (UniProt accession #Q5ZPR3 - isoform 1) and a C-terminal Fc fusion from human IgG1, which exists as a dimer under non-reducing condition 

 

Source:  Human cells stably expressing B7-H3b-Fc and growing in chemical-defined media with no animal components or antibiotics

 

Deduced Amino Acid Sequence:

        10         20         30         40         50         60 

LEVQVPEDPV VALVGTDATL CCSFSPEPGF SLAQLNLIWQ LTDTKQLVHS FAEGQDQGSA 

        70         80         90        100        110        120 

YANRTALFPD LLAQGNASLR LQRVRVADEG SFTCFVSIRD FGSAAVSLQV AAPYSKPSMT 

       130        140        150        160        170        180 

LEPNKDLRPG DTVTITCSSY QGYPEAEVFW QDGQGVPLTG NVTTSQMANE QGLFDVHSIL 

       190        200        210        220        230        240 

RVVLGANGTY SCLVRNPVLQ QDAHSSVTIT PQRSPTGAVE VQVPEDPVVA LVGTDATLRC 

       250        260        270        280        290        300 

SFSPEPGFSL AQLNLIWQLT DTKQLVHSFT EGRDQGSAYA NRTALFPDLL AQGNASLRLQ 

       310        320        330        340        350        360 

RVRVADEGSF TCFVSIRDFG SAAVSLQVAA PYSKPSMTLE PNKDLRPGDT VTITCSSYRG 

       370        380        390        400        410        420 

YPEAEVFWQD GQGVPLTGNV TTSQMANEQG LFDVHSVLRV VLGANGTYSC LVRNPVLQQD 

       430        440        450        460        470        480 

AHGSVTITGQ PMTFPPEAST GTHTCPPCPA PELLGGPSVF LFPPKPKDTL MISRTPEVTC 

       490        500        510        520        530        540 

VVVDVSHEDP EVKFNWYVDG VEVHNAKTKP REEQYNSTYR VVSVLTVLHQ DWLNGKEYKC 

       550        560        570        580        590        600 

KVSNKALPAP IEKTISKAKG QPREPQVYTL PPSREEMTKN QVSLTCLVKG FYPSDIAVEW 

       610        620        630        640        650        660 

ESNGQPENNY KTTPPVLDSD GSFFLYSKLT VDKSRWQQGN VFSCSVMHEA LHNHYTQKSL 

 

SLSPGK 

 

M.W.:  Calculated molecular mass 72.6 kDa; estimated by SDS-PAGE under reducing condition ~95 kDa and ~65 kDa probably representing the 4Ig-B7-H3 and 2Ig-B7-H3 isoforms

 

Calculated PI: 5.29

 

Calculated Extinction Coefficients (M-1 cm-1, at 280nm): 73185

 

Purity:>95% judged by SDS-PAGE under reducing condition with two major products: 25% of 95 kDa and 75% of 65 kDa (see the gel image above)

 

Formulation: Supplied at 0.5 mg/ml in sterile PBS pH7.4 (carrier free).

 

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

 

Biological Activity: Inhibits anti­CD3­induced proliferation of stimulated human T cells.  Binds anti-B7-H3 monoclonal antibodies, human IgG1 (SKU#MAB1755) and rabbit IgG (SKU#MAB1753) in a functional ELISA (see Technical Data) 

  

 

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.

 

 

Technical Data

 

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.

 

Storage

 

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.

 

References

 

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.

 

Documentation

 

Product datasheet (pdf) can be downloaded here: FCL1662-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.