B7-H3/CD276 Antibody [Unconjugated]

B7-H3 was detected in immersion fixed paraffin-embedded sections of human melanoma using Goat Anti-Human B7-H3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1027) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Western blot shows lysates of U2OS human osteosarcoma parental cell line and B7-H3 knockout U2OS cell line (KO). PVDF membrane was probed with 1 µg/mL of Goat Anti-Human B7‑H3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1027) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for B7‑H3 at approximately 95 kDa (as indicated) in the parental U2OS cell line, but is not detectable in knockout U2OS cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

PC-3 human prostate carcinoma cells were stained with Goat Anti-Human B7‑H3 Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF1027, filled histogram) or control antibody (Catalog #AB‑108‑C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107).

Western blot shows lysates of LNCaP human prostate cancer cell line and U2OS human osteosarcoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human B7‑H3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1027) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for B7‑H3 at approximately 90-110 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Simple Western lane view shows lysates of U2OS human osteosarcoma cell line, loaded at 0.2 mg/mL. A specific band was detected for B7‑H3 at approximately120-160 kDa (as indicated) using 10 µg/mL of Goat Anti-Human B7‑H3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1027) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

• Reactivity: Hu
• Applications: WB, Simple Western, Flow, IHC, CyTOF-ready, KO
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

B7-H3/CD276 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human B7‑H3
  Leu29-Pro245
  Accession # NP_079516
• Specificity: Detects human B7-H3 in direct ELISAs and Western blots. In direct ELISAs, approximately 5% cross-reactivity with recombinant human (rh) B7‑H2 is observed, and less than 1% cross-reactivity with rhB7-1, rhB7-2, rhB7-H1, and rhPD-L2 is observed.
• Details of Functionality: ActivityAssaywCitation not found None None None None None
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: CD276

Applications/Dilutions

• Dilutions:
  • Western Blot 1 ug/mL
  • Simple Western 10 ug/mL
  • Flow Cytometry 0.25 ug/10^6 cells
  • Immunohistochemistry 5-15 ug/mL
  • CyTOF-ready
  • Knockout Validated
• Control:

  B7-H3/CD276 Knockout HeLa Cell Lysate

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for B7-H3/CD276 Antibody [Unconjugated]

• B7H3
• B7-H3
• B7H34Ig-B7-H3
• B7-H3B7 homolog 3
• CD276 antigen
• CD276 molecule
• CD276
• Costimulatory molecule

Background

Human B7 homolog 3 (B7-H3) is a member of the B7 family of immune proteins that provide signals for regulating immune responses (1‑3). Other family members include B7-1, B7-2, B7-H2, PD-L1 (B7-H1), and PD-L2. B7 proteins are immunoglobulin (Ig) superfamily members with extracellular Ig-V-like and Ig-C-like domains and short cytoplasmic domains. Among the family members, they share about 20‑40% amino acid (aa) sequence identity. The cloned human B7-H3 cDNA encodes a 316 aa type I membrane precursor protein with a putative 28 aa signal peptide, a 217 aa extracellular region containing one V-like and one C-like Ig domain, a transmembrane region, and a 45 aa cytoplasmic domain. An isoform of human B7-H3 containing a four-Ig-like domain extracellular region has also been identified. Human B7-H3 is not expressed on resting B cells, T cells, monocytes or dendritic cells, but is induced on dendritic cells and monocytes by inflammatory cytokines. B7-H3 expression is also detected on various normal tissues and in some tumor cell lines. Human B7-H3 does not bind any known members of the CD28 family of immunoreceptors. However, B7-H3 has been shown to bind an unidentified counter-receptor on activated T cells to costimulate the proliferation of CD4⁺ or CD8⁺ T cells. B7-H3 has also been found to enhance the induction of primary cytotoxic T lymphocytes and stimulate IFN-gamma production (1‑3).

1. Chapoval, A.I. et al. (2001) Nat. Immunol. 2:269.
2. Sharpe, A.H. and G.J. Freeman (2002) Nat. Rev. Immunol. 2:116.
3. Coyle, A. and J. Gutierrez-Ramos (2001) Nat. Immunol. 2:203.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for B7-H3/CD276 Antibody (AF1027)(12)

Publications using AF1027

• T Shi, Y Ma, L Cao, S Zhan, Y Xu, F Fu, C Liu, G Zhang, Z Wang, R Wang, H Lu, B Lu, W Chen, X Zhang B7-H3 promotes aerobic glycolysis and chemoresistance in colorectal cancer cells by regulating HK2 Cell Death Dis, 2019;10(4):308. 2019 [PMID: 30952834] (Western Blot, Human)
• J Zhang, J Wang, DM Marzese, X Wang, Z Yang, C Li, H Zhang, J Zhang, CC Chen, DF Kelly, W Hua, DSB Hoon, Y Mao B7H3 regulates differentiation and serves as a potential biomarker and theranostic target for human glioblastoma Lab. Invest., 2019;0(0):. 2019 [PMID: 30914782] (Western Blot, Human)
• KE Wilson, SV Bachawal, L Abou-Elkac, K Jensen, S Machtaler, L Tian, JK Willmann Spectroscopic Photoacoustic Molecular Imaging of Breast Cancer using a B7-H3-targeted ICG Contrast Agent Theranostics, 2017;7(6):1463-1476. 2017 [PMID: 28529630] (IHC, Human)
• D Quandt, E Fiedler, A Müller, WC Marsch, B Seliger High constitutive B7-H3 expression on human keratinocytes supports T cell immunity J. Dermatol. Sci., 2017;0(0):. 2017 [PMID: 28465042] (IHC, Human)
• Y Li, G Guo, J Song, Z Cai, J Yang, Z Chen, Y Wang, Y Huang, Q Gao B7-H3 Promotes the Migration and Invasion of Human Bladder Cancer Cells via the PI3K/Akt/STAT3 Signaling Pathway J Cancer, 2017;8(5):816-824. 2017 [PMID: 28382144] (IHC-P, Human)
• BR Yoon, YH Chung, SJ Yoo, K Kawara, J Kim, IS Yoo, CG Park, SW Kang, WW Lee Preferential Induction of the T Cell Auxiliary Signaling Molecule B7-H3 on Synovial Monocytes in Rheumatoid Arthritis J. Biol. Chem, 2016;291(8):4048-57. 2016 [PMID: 26702052] (Western Blot, Human)
• Bachawal S, Jensen K, Wilson K, Tian L, Lutz A, Willmann J Breast Cancer Detection by B7-H3-Targeted Ultrasound Molecular Imaging. Cancer Res, 2015;75(12):2501-9. 2015 [PMID: 25899053] (IHC-P, Human)
• Sun J, Fu F, Gu W, Yan R, Zhang G, Shen Z, Zhou Y, Wang H, Shen B, Zhang X Origination of new immunological functions in the costimulatory molecule B7-H3: the role of exon duplication in evolution of the immune system. PLoS ONE, 2011;6(9):e24751. 2011 [PMID: 21931843] (Western Blot, Human)
• Nagai Y, Aso H, Ogasawara H, Tanaka S, Taketa Y, Watanabe K, Ohwada S, Rose MT, Kitazawa H, Yamaguchi T Anterior pituitary progenitor cells express costimulatory molecule 4Ig-B7-H3. J. Immunol., 2008;181(9):6073-81. 2008 [PMID: 18941196] (Western Blot, Bovine)
• Tran CN, Thacker SG, Louie DM, Oliver J, White PT, Endres JL, Urquhart AG, Chung KC, Fox DA Interactions of T cells with fibroblast-like synoviocytes: role of the B7 family costimulatory ligand B7-H3. J. Immunol., 2008;180(5):2989-98. 2008 [PMID: 18292521] (ICC, Human)
• Jang BC, Park YK, Choi IH, Kim SP, Hwang JB, Baek WK, Suh MH, Mun KC, Suh SI 12-O-tetradecanoyl phorbol 13-acetate induces the expression of B7-DC, -H1, -H2, and -H3 in K562 cells. Int. J. Oncol., 2007;31(6):1439-47. 2007 [PMID: 17982670] (Flow Cytometry, Human)
• Petroff MG, Kharatyan E, Torry DS, Holets L The immunomodulatory proteins B7-DC, B7-H2, and B7-H3 are differentially expressed across gestation in the human placenta. Am. J. Pathol., 2005;167(2):465-73. 2005 [PMID: 16049332] (Western Blot, Human)

Review for B7-H3/CD276 Antibody (AF1027) (1)

Average Rating: 4

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Anonymous IHC-Fr Human 01/05/2015

Summary

• Application: Immunohistochemistry-Frozen
• Sample Tested: See PMID 22080438
• Species: Human

Bioinformatics

• Gene Symbol: CD276
• Uniprot:
  • NP_079516()