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Anti-B7-H3 (CD276), clone 376.96, Cat. No. MABC1731, is a mouse monoclonal antibody that detects human B7-H3 (CD276) and has been tested for use in Flow Cytometry, Inhibition/Function Analysis, Immunoprecipitation, and Western Blotting.
More>>Anti-B7-H3 (CD276), clone 376.96, Cat. No. MABC1731, is a mouse monoclonal antibody that detects human B7-H3 (CD276) and has been tested for use in Flow Cytometry, Inhibition/Function Analysis, Immunoprecipitation, and Western Blotting. Less<<
CD276 antigen (UniProt: Q5ZPR3; also known as 4Ig-B7-H3, B7 homolog 3, B7-H3, Costimulatory molecule, CD276) is encoded by the CD276 (also known as B7H3, PSEC0249, UNQ309/PRO352) gene (Gene ID: 80381) in human. CD276 is a single-pass type I membrane protein that is member of B7 immunoregulatory family. It participates in the regulation of T-cell-mediated immune response. It is a ubiquitous protein but is undetectable in peripheral blood lymphocytes or granulocytes and is only weakly expressed in resting monocytes. It is also expressed in dendritic cells derived from monocytes and in epithelial cells of sinonasal tissue, extravillous trophoblast cells and Hofbauer cells of the first trimester placenta and term placenta. It plays a key role in providing the placenta and fetus with a suitable immunological environment throughout pregnancy. Higher expression of CD276 has been shown in a variety of cancer types, including breast, pancreatic, and ovarian cancer and its expression has been correlated with poor survival in ovarian cancer subjects. It is suggested to play a protective role in tumor cells by inhibiting natural-killer mediated cell lysis. Its levels are also reported to be up-regulated in cells that mediate rejection of human transplants. CD276 is synthesized with a signal peptide (aa 1-28), which is subsequently cleaved off to produce the mature form, which contains an extracellular domain (aa 29-466), a short transmembrane domain (aa 467-487), and a cytoplasmic domain (aa 488-534). Clone 376.96 is reported to inhibit the growth of chemoresistant ovarian cancer cells in vitro. (Ref.: Fauci, JM., et al. (2014). Gynecologic Oncology 132(1); 203-210; Chen, Y-W., et al. (2008). Curr. Can. Drug Targets. 8(5); 404-413).
References
Product Information
Format
Purified
Presentation
Purified mouse monoclonal antibody IgG2a in PBS without azide.
Applications
Application
Anti-B7-H3 (CD276), clone 376.96, Cat. No. MABC1731, is a mouse monoclonal antibody that detects human B7-H3 (CD276) and has been tested for use in Flow Cytometry, Inhibition/Function Analysis, Immunoprecipitation, and Western Blotting.
Key Applications
Flow Cytometry
Inhibits Activity/Function
Immunoprecipitation
Western Blotting
Application Notes
Western Blotting Analysis: A representative lot detected B7-H3 (CD276) in Western Blotting applications (Chen, Y.W., et. al. (2008). Curr Cancer Drug Targets. 8(5):404-13).
Immunoprecipitation Analysis: A representative immunoprecipitated B7-H3 (CD276) in Immunoprecipitation applications (Chen, Y.W., et. al. (2008). Curr Cancer Drug Targets. 8(5):404-13).
Inhibition/Function Analysis: A representative lot inhibited the growth of chemoresistant ovarian cancer cells in vitro (Chen, Y.W., et. al. (2008). Curr Cancer Drug Targets. 8(5):404-13; Fauci, J.M., et. al. (2014). Gynecol Oncol. 132(1):203-10).
Flow Cytometry Analysis: A representative lot detected B7-H3 (CD276) in Flow Cytometry applications (Chen, Y.W., et. al. (2008). Curr Cancer Drug Targets. 8(5):404-13; Fauci, J.M., et. al. (2014). Gynecol Oncol. 132(1):203-10).
Biological Information
Immunogen
Cultured COLO 38 human melanoma cells.
Epitope
extracellular domain
Clone
376.96
Concentration
Please refer to lot specific datasheet.
Host
Mouse
Specificity
Clone 376.96 is a mouse monoclonal antibody that specifically detects human B7-H3 (CD276). It targets an epitope within the extracellular domain.
Flow Cytometry Analysis: 1 µg of this antibody detected B7-H3 (CD276) in one million SK-OV-3 cells.
Usage Statement
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage Conditions
Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
The high rate of relapse in patients with advanced ovarian cancer likely reflects the chemoresistance of cancer initiating cells (CICs). We evaluated the anti-tumor activity of monoclonal antibody (mAb) 376.96, which recognizes a B7-H3 epitope expressed on ovarian carcinoma cells (OCCs), in combination with the tyrosine kinase inhibitor Sunitinib and chemotherapy on chemosensitive and chemoresistant cells and CICs.Eight ovarian cancer cell lines including platinum- and taxane-resistant cell lines were analyzed by flow cytometry to establish expression of the mAb 376.96-defined-B7-H3-epitope on differentiated ovarian cancer cells and CICs. Samples from 10 ovarian cancer patients were analyzed via immunohistochemistry for mAb 376.96-defined-B7-H3-epitope expression. In vitro studies assessed mAb 376.96 alone and in combination with Sunitinib on the growth of chemosensitive and chemoresistant cell lines and on the content of CICs.The mAb-376.96-defined-B7-H3 epitope is expressed on both differentiated cells and CICs in chemosensitive and chemoresistant ovarian cancer cell lines and 10 patient derived ovarian cancer tumors. In vitro treatment of chemoresistant cell lines with mAb 376.96 resulted in decreased cell viability. mAb 376.96 enhanced the cytotoxicity of Sunitinib and reduced the content of CICs.The mAb-376.96-defined-B7-H3-epitope was found to be expressed on both differentiated ovarian cancer cells and CICs in chemosensitive and chemoresistant ovarian cancer cell lines. mAb 376.96 inhibited the in vitro growth of chemosensitive and chemoresistant OCCs and reduced the content of CICs when used with Sunitinib. Further studies examining B7-H3 as a potential target of mAb-based immunotherapy for this type of malignancy are warranted.
The immunoregulatory protein human B7H3 is a tumor-associated antigen that regulates tumor cell migration and invasion. Chen, YW; Tekle, C; Fodstad, O Curr Cancer Drug Targets
8
404-13
2008
The monoclonal antibody (mAb) 376.96 has been used for detection of micrometastatic tumor cells due to its high binding specificity for a wide range of tumor cells, but the identity and function of its target antigen have not been known. Here, using immunoprecipitation and siRNA technology, we demonstrate that the antigen is the human 4Ig-B7H3 (4Ig-hB7H3) protein, previously known as an immunoregulatory protein in immune cells. Immunoblots of whole cell lysates, subcellular fractionation and tunicamycin treatment of human tumor cells indicated that 4Ig-hB7H3 is a approximately 100-kDa N-linked glycosylated membrane protein. The tumor promoter phorbol 12-myristate 13-acetate (PMA) enhanced the expression of 4Ig-hB7H3 in FEMX-I (melanoma), MA11 (breast cancer), and OHS (osteosarcoma) cells, suggesting that 4Ig-hB7H3 may be implicated in tumorigenesis. Most importantly, siRNA-downregulation of hB7H3 reduced cell adhesion to fibronectin of melanoma and breast cancer cells by up to 50 %, and migration and matrigel-invasion by more than 70 %, but surprisingly had no apparent impact on cell proliferation. In conclusion, our data present 4Ig-hB7H3 as a tumor-associated antigen and suggests a novel biological role of 4Ig-hB7H3 in tumor progression and metastasis.
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