Recombinant Human CD276 antigen(CD276),partial (Active)

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Code CSB-MP733578HU
Size US$298Purchase it in Cusabio online store
(only available for customers from the US)
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  • (Tris-Glycine gel) Discontinuous SDS-PAGE (reduced) with 5% enrichment gel and 15% separation gel.

  • Activity
    Measured by its binding ability in a functional ELISA. Immobilized CD276 at 2 μg/ml can bind Anti-CD276 rabbit monoclonal antibody, the EC50 of human CD276 protein is 1.961-2.243 ng/ml. Biological Activity Assay

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Product Details

Description

The DNA fragment corresponding to Leu29-Gly245 of the human CD276 antigen is fused with an FC-Myc-tag at the C-terminus and then is expressed in the mammalian cells. The resulting product is the recombinant human CD276 antigen. Its purity is over 93% measured by SDS-PAGE. On the gel, this protein was migrated to the molecular weight band of about 60-80 kDa. The endotoxin content of this CD276 protein is less than 1.0 EU/ug determined by the LAL method. Its bioactivity has been validated through an ELISA. In the functional ELISA, this recombinant CD276 protein can bind to the anti-CD276 antibody, and the EC50 is 1.961-2.243 ng/ml. This CD276 protein is available now.

CD276 is an immune checkpoint molecule in the epithelial-mesenchymal transition (EMT) pathway. It is involved in cell proliferation, invasion, and metastasis in malignancies. Overexpression of CD276 has been found in numerous solid tumors such as breast, colorectal, and lung cancer, and has been strongly linked to a poor clinical prognosis.

Purity Greater than 93% as determined by SDS-PAGE.
Endotoxin Less than 1.0 EU/ug as determined by LAL method.
Activity Measured by its binding ability in a functional ELISA. Immobilized CD276 at 2 μg/ml can bind Anti-CD276 rabbit monoclonal antibody, the EC50 of human CD276 protein is 1.961-2.243 ng/ml.
Target Names CD276
Uniprot No. Q5ZPR3
Research Area Cancer
Alternative Names 4Ig B7 H3; 4Ig-B7-H3; AU016588; B7 H3; B7 homolog 3; B7-H3; B7H3; B7RP-2; CD_antigen=CD276; CD276; CD276 antigen; CD276 molecule; CD276_HUMAN; Costimulatory molecule; Flags: Precursor; PSEC0249; UNQ309/PRO352
Molecular Characterization
Species Homo sapiens (Human)
Source Mammalian cell
Expression Region 29-245aa
Target Protein Sequence LEVQVPEDPVVALVGTDATLCCSFSPEPGFSLAQLNLIWQLTDTKQLVHSFAEGQDQGSAYANRTALFPDLLAQGNASLRLQRVRVADEGSFTCFVSIRDFGSAAVSLQVAAPYSKPSMTLEPNKDLRPGDTVTITCSSYQGYPEAEVFWQDGQGVPLTGNVTTSQMANEQGLFDVHSILRVVLGANGTYSCLVRNPVLQQDAHSSVTITPQRSPTG
Mol. Weight 53.4 kDa
Protein Length Partial
Tag Info C-terminal FC-Myc-tagged
Form Liquid or Lyophilized powder
Buffer Lyophilized from a 0.2 μm filtered PBS, 6% Trehalose, pH 7.4
Troubleshooting
and FAQs
Protein FAQs
Storage Condition Store at -20°C upon receipt, aliquoting is necessary for mutiple use. Avoid repeated freeze-thaw cycles.
Shelf Life The shelf life is related to many factors, storage state, buffer ingredients, storage temperature and the stability of the protein itself.
Generally, the shelf life of liquid form is 6 months at -20°C/-80°C. The shelf life of lyophilized form is 12 months at -20°C/-80°C.
Lead Time Basically, we can dispatch the products out in 1-3 working days after receiving your orders. Delivery time may differ from different purchasing way or location, please kindly consult your local distributors for specific delivery time.
Note: All of our proteins are default shipped with normal blue ice packs, if you request to ship with dry ice, please communicate with us in advance and extra fees will be charged.
Notes Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.
Datasheet & COA Please contact us to get it.

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 Q&A
Q:

Are CUSABIO recombinant proteins sterile?

A:

Yes, we can offer an aseptic processing service and it is free of charge, but you should remark this information when placing the order. We've performed aseptic processing for liquid protein before lyophilization, but there may exist contamination during lyophilization process. To clarify, lyophilized proteins can’t guarantee aseptic processing for the whole manufacture process.

Q:

Can you remove the endotoxin?

A:

Not all endotoxin can be removed. Please contact us in advance if you need to remove the tag which takes 2-3 business days. We could offer endotoxin removal service free of charge using PMB affinity chromatography, use LAL reagent to semi-quantitatively detect the content of endotoxin, and guarantee endotoxin level within 0.1 ng/μg (1 EU/μg).

Q:

How to avoid inclusion bodies and improve soluble expression?

A:

a. For proteins with high hydrophobicity or transmembrane domains, you should add fusion tags or add heat shock chaperones, and induce for a shorter time at low temperature or change to poor media. Generate truncated forms of protein or use membrane-rich strains.
b. For incorrect disulfide bond formation, you should add fusion partners, including thioredoxin, DsbA, DsbC. Clone in a vector containing secretion signal peptide to cell periplasm. Use gamiB (DE3)strains with the oxidative cytoplasmic environment. Lower inducer concentration and induction temperature.
c. For incorrect folding, you should use a fusion partner. Co-express with molecular chaperones. Use strains with cold-adapted chaperones. Supplement media with chemical chaperones and cofactors. Reduce the inducer concentration and add fresh media. Induce for a shorter time at low temperature.

Target Background

Function
(From Uniprot)

CD276 (B7-H3) signal pathway and function
Early studies found that CD276 can promote the activation of T cells. Chapoval et al. confirmed that in the presence of anti-CD3 antibodies, CD276 can promote the proliferation of CD4 and CD8+ T cells and selectively promote the secretion of IFN-γ. Moreover, the transfection of CD276 into tumor cells will enhance the killing ability of CTL. Further research found that only TLT-2 transgenic cells can bind to mouse CD276 with high affinity, confirming that TLT-2 is a receptor molecule for CD276. And Hashiguchi et al. confirmed that the CD276-TLT-2 pathway can enhance the activation of T cells. However, Leitner et al. used flow cytometry to find no specific binding between CD276 and TLT-2. Therefore, the exact receptor molecule of CD276 is still unclear.
On the other hand, studies have found that CD276 can also suppress T cell immune responses. Early studies have shown that CD276 can inhibit the activation of human and mouse T cells by activating or inhibiting NFTA (nuclear factor for activated T cells), NF-KB (nuclear factor kB), and AP-1 (activator protein-1) pathways. In addition, there are results that prove that CD276 may inhibit T cell immune response by inhibiting the activity of Th1. The study by Leiner et al. also found that CD276 can down-regulate the secretion of IL-2 from T cells, thereby inhibiting the activity of T cells.

CD276 (B7-H3) as a target in tumor treatment
B7-H3 also plays a key role in a variety of tumor processes. Tumor patients with high expression of B7-H3 are often accompanied by postoperative tumor proliferation, high tumor recurrence rate, and tumor-induced death. Most studies have confirmed that B7-H3 plays a role in suppressing immune response in tumor immunity. A few studies have confirmed that B7-H3 also participates in the positive immune response to tumor cells.
① High expression of B7-H3 in tumor tissues can promote tumor occurrence and metastasis.
②B7-H3 can inhibit T cell-mediated cellular immunity, allowing tumors to escape immune surveillance.
③B7-H3 is involved in the drug resistance of tumor cells.
④B7-H3 may be involved in tumor angiogenesis.
⑤ The expression of B7-H3 is negatively correlated with tumor progression. Studies have found that patients expressing B7-H3 have a better prognosis. Loos et al. found that B7-H3 inhibits tumor progression in pancreatic cancer, which may be achieved by promoting the secretion of IFN-γ. However, in the study of gastric cancer, Arigami et al. found that patients with more expression of B7-H3 in the circulation, the more severe the pathological process of their tumors, and the worse the prognosis. And Yamato et al.'s research in pancreatic cancer showed that the high expression of B7-H3 is related to the more advanced pathological process of the tumor and lymph node metastasis. Relevant evidence shows that the relationship between tumors and B7-H3 shows a certain degree of complexity whether on different tumors or on the same tumor.

Gene References into Functions
  1. High B7-H3 expression is associated with phyllodes tumors. PMID: 30486739
  2. characterization of the expression pattern and biological function of B7-H3 in brain gliomas; B7-H3 expression is regulated by multiple mechanisms and is potentially involved in the T-cell receptor signaling pathway; higher B7-H3 expression indicates a worse prognosis for glioma patients PMID: 30027617
  3. High expression level of B7H3 in muscleinvasive bladder cancer (MIBC) tissues is associated with a poor clinico-pathological status and poor prognosis, and promotes the development of MIBC in vitro and in vivo. PMID: 30132557
  4. Tumor B7-H3 expression was significantly higher in immunocompetent vs. immunosuppressed patients, largely driven by very low expression in HIV+ patients. PMID: 29484464
  5. Colorectal cancer (CRC) patients expressing both high B7-H3 and high B3GALT4 contributed to a significant decrease in overall survival. The expression of B3GALT4 in CRC is positively correlated with B7-H3 expression in vitro. B7-H3/B3GLAT4 may be used as dual prognostic biomarkers for CRC. PMID: 30131660
  6. this study shows reduced sB7-H3 expression in the peripheral blood of systemic lupus erythematosus patients PMID: 29423417
  7. High B7-H3 expression is associated with cervical cancer invasiveness. PMID: 28627681
  8. IDH1 and B7H3 cannot be used as independent prognostic factors, co-expression of IDH1 and B7H3 significantly correlated with the prognosis of CRC patients and may serve as a combined predictive marker. Thus, the correlation between IDH1 and B7H3 has been proven in vivo and in vitro. PMID: 29871819
  9. Our findings revealed that B7-H3 affect ovarian cancer progression through the Jak2/Stat3 pathway, indicating that B7-H3 has the potential to be a useful prognostic marker. PMID: 28765941
  10. metastatic melanoma cells with knockdown expression of B7-H3 showed modest decrease in proliferation and glycolytic capacity. PMID: 28513992
  11. results suggest that B7-H3 may be a valuable biomarker in determining tumor progression and prognosis of intrahepatic cholangiocarcinoma PMID: 29696716
  12. these results demonstrate that sB7-H3 promotes invasion and metastasis through the TLR4/NF-kappaB pathway in pancreatic carcinoma PMID: 27273624
  13. Inhibition of the B7-H3 immune checkpoint limits tumor growth by enhancing cytotoxic lymphocyte function PMID: 28685773
  14. Higher B7-H3 expression correlates with Gleason grade, prostate cancer stage and poor oncologic outcomes in prostatectomy cohorts. B7-H3 expression appears to be related to androgen signaling as well as the immune reactome. PMID: 27801901
  15. Review/Meta-analysis: High B7-H3 expression was a significant indicator of lymph node metastasis and advanced TNM stage in non-small cell lung cancer. PMID: 27835582
  16. B7-H3 promotes epithelial mesenchymal transformation in colorectal cancer cells by activating the PI3K-Akt pathway and upregulating the expression of Smad1. PMID: 27145365
  17. Plasma B7-H3 levels were decreased significantly in children subjected to pediatric general and cardiac surgery, which is closely associated with the severity of surgical stress. The negative correlation of plasma B7-H3 levels at day 1 and day 3 after surgery with surgical stress scoring implicates that the plasma B7-H3 level might be a useful biomarker for monitoring stress intensity during pediatric surgery. PMID: 27459969
  18. B7-H3 promotes the oxaliplatin resistance in colorectal cancer cells upregulating the expression of XRCC1 via PI3K-AKT pathway. PMID: 28676400
  19. Children with Mycoplasma pneumoniae pneumonia had significantly higher levels of sB7-H3 and IL-36 compared to control subjects. PMID: 27188891
  20. CD276 is broadly expressed by tumor cells and tumor vasculature but Is dispensable for tumor growth. PMID: 28399408
  21. B7-H3 hijacks SREBP-1/FASN signaling mediating abnormal lipid metabolism in lung cancer PMID: 27939887
  22. We found that B7-H3 promoted the Warburg effect, evidenced by increased glucose uptake and lactate production in B7-H3-expressing cells. B7-H3 also increased the protein levels of HIF1alpha and its downstream targets, LDHA and PDK1, key enzymes in the glycolytic pathway PMID: 27197253
  23. Upregulated sB7-H3 expression in MPEs is correlated with TNM stage of NSCLC and may serve as a potential biomarker for NSCLC-derived MPEs PMID: 27071700
  24. High expression of B7-H3 is associated with Colorectal cancer. PMID: 26787540
  25. Elevated B7-H3 expression is significantly associated with poor survival in cancer patients. (Meta-analysis) PMID: 27626927
  26. The results provide novel insights into the function of B7-H3 in cancer, and suggest that targeting of B7-H3 may be a novel alternative to improve current anticancer therapies. PMID: 26771843
  27. Suggesting that B7-H3 and Tregs may act cooperatively in tumor immune evasion, leading to poor outcomes for NSCLC patients. PMID: 26823710
  28. B7-H3 is one of the most strongly expressed B7-family molecules in AML and merits further investigation. PMID: 26376842
  29. findings demonstrate that activation-induced B7-H3 expression on synovial monocytes has the potential to inhibit Th1-mediated immune responses and immunomodulatory roles affecting RA pathogenesis. PMID: 26702052
  30. B7H3 promotes cell migration and invasion through the Jak2/Stat3/MMP9 signaling pathway in colorectal cancer PMID: 26151358
  31. B7-H1 and B7-H3 are independent predictors of poorer survival in patients with non-small cell lung cancer. PMID: 25609202
  32. B7-H3 may play an important role in asthma exacerbation and was a useful clinical biomarker to evaluate asthma exacerbation. PMID: 26108069
  33. found that the N-glycans of B7-H3 from Ca9-22 oral cancer cells contain the terminal alpha-galactose and are more diverse with higher fucosylation and better interaction with DC-SIGN and Langerin on immune cells than that from normal cells PMID: 26438868
  34. Study shows that B7-H3 promotes mantle cell lymphoma progression and its knockdown significantly enhances the chemosensitivity. PMID: 25872657
  35. These results demonstrated that B7-H3 expression in acute leukemia predicts an unfavorable outcome. PMID: 25130683
  36. Our results indicate that B7-H3 expression in cervical invasive squamous cell carcinoma may play an important role in overcoming CD8(+) T-cell immunoregulation to acquire aggressive growth. PMID: 25675190
  37. Data shows that B7-H3 is aberrantly expressed in gastric cancer. In addition to modulating tumor immunity, B7-H3 may have a novel role in regulating SGC-7901 cell metastasis. PMID: 25120098
  38. Overexpression of B7-H3 in CD14+ monocytes is associated with renal cell carcinoma progression.B7-H3 might play an important role in angiogenesis of renal cell carcinoma mediated by CD14(+) monocytes. PMID: 25416051
  39. Data show that immunoglobulin-like transcript 4 (ILT4) increases the expression of the co-inhibitory molecule B7-H3 through PI3K/AKT/mTOR signalling. PMID: 26149216
  40. IL-2/IL-2R and B7-H3 pathways may be involved in the progression of clear cell renal cell carcinoma PMID: 25089268
  41. The overexpression of B7-H3 induces resistance to apoptosis in colorectal cancer cell lines by upregulating the Jak2-STAT3 signaling pathway. PMID: 25684945
  42. B7-H3 was significantly up-regulated on monocytes in rheumatoid arthritis patients, while soluble B7-H3 in serum was decreased. A polymorphism variant, B7-H3-T-A-C-T, was shown to be associated with the incidence of RA and the decreased release of sB7-H3. PMID: 25931383
  43. B7-H3-mediated STAT3 signaling pathway is an important mechanism for inducing M2-type polarization of tumor associated macrophages. PMID: 25370943
  44. The study confirms the overexpression of B7-H3 in colorectal cancer and shows the main localization of the protein in cytoplasm and cell membrane. PMID: 25139714
  45. High level of serum B7-H3 in patients with Hepatocellular Carcinoma is caused by the increased expression of a newly discovered spliced soluble B7-H3 isoform in carcinoma and peritumor tissues. PMID: 24194851
  46. Data indicate that six sites of co-stimulatory molecule B7-H3 gene have single nucleotide polymorphisms variations and five sites are related to the pathogenesis of rheumatoid arthritis (RA). PMID: 25200161
  47. Study shows that CD276 can be used to discriminate ECs from malignant tissue from ECs from normal tissue. In addition, CD276(+) CEC do occur in higher frequencies in patients with advanced cancer. PMID: 24892449
  48. Data indicate that the expression of membrane B7-H3 (mB7-H3) in peri-tumor normal tissues was higher than that in tumor tissues, and it can serve as an assistant diagnostic marker for hepatitis B virus-associated hepatocellular carcinoma (HBV-HCC). PMID: 25001942
  49. B7H3 and ATP1B3 are overexpressed in tumor endothelial cells, favoring an angiogenic phenotype. PMID: 24236063
  50. Thus, B7-H3 may have a critical role in primary hepatocellular carcinoma and it may enhance tumor escape from the immune surveillance of CD8(+) T cells. PMID: 24787022

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Subcellular Location Membrane, Single-pass type I membrane protein
Protein Families Immunoglobulin superfamily, BTN/MOG family
Tissue Specificity Ubiquitous but not detectable in peripheral blood lymphocytes or granulocytes. Weakly expressed in resting monocytes. Expressed in dendritic cells derived from monocytes. Expressed in epithelial cells of sinonasal tissue. Expressed in extravillous trophob
Database Links

HGNC: 19137

OMIM: 605715

KEGG: hsa:80381

STRING: 9606.ENSP00000320084

UniGene: Hs.744915

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