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Human B7-H3 Antibody

Catalog #: AF1027
40 Citations 1 Review Datasheet / COA / SDS
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AF1027
AF1027-SP
Detection of Human B7‑H3 by Western Blot.
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Citations (40)
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Reviews (1)
Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human B7-H3 Antibody Summary

Species Reactivity
Human
Specificity
Detects human B7-H3 in direct ELISAs and Western blots.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant human B7‑H3
Leu29-Pro245
Accession # NP_079516
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Simple Western
10 µg/mL
See below
Flow Cytometry
0.25 µg/106 cells
See below
Immunohistochemistry
5-15 µg/mL
See below
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 
Knockout Validated
B7‑H3 is specifically detected in U2OS human osteosarcoma parental cell line but is not detectable in B7‑H3 knockout U2OS cell line.
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human B7-H3 antibody by Western Blot. View Larger

Detection of Human B7‑H3 by Western Blot. 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.

Flow Cytometry Detection of B7‑H3 antibody in PC‑3 Human Cell Line antibody by Flow Cytometry. View Larger

Detection of B7‑H3 in PC‑3 Human Cell Line by Flow Cytometry. 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).

Immunohistochemistry B7-H3 antibody in Human Melanoma by Immunohistochemistry (IHC-P). View Larger

B7-H3 in Human Melanoma. 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.

Simple Western Detection of Human B7-H3 antibody by Simple Western<sup>TM</sup>. View Larger

Detection of Human B7‑H3 by Simple WesternTM. 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 approximately 120-160 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.

Knockout Validated Western Blot Shows Human B7-H3 Antibody Specificity by Using Knockout Cell Line. View Larger

Western Blot Shows Human B7‑H3 Specificity by Using Knockout Cell Line. 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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & 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.

Background: B7-H3

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).

References
  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.
Long Name
B7 Homolog 3
Entrez Gene IDs
80381 (Human); 102657 (Mouse); 315716 (Rat); 102131295 (Cynomolgus Monkey)
Alternate Names
B7H3; B7-H3; B7H34Ig-B7-H3; B7-H3B7 homolog 3; CD276 antigen; CD276 molecule; CD276; Costimulatory molecule

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Citations for Human B7-H3 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

40 Citations: Showing 1 - 10
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  1. Prognostic value of B7-H1, B7-H3 and the stage, size, grade and necrosis (SSIGN) score in metastatic clear cell renal cell carcinoma
    Authors: Mischinger, J;Frohlich, E;Mannweiler, S;Meindl, C;Absenger-Novak, M;Hutterer, GC;Seles, M;Augustin, H;Chromecki, TF;Jesche-Chromecki, J;Pummer, K;Zigeuner, R;
    Cent European J Urol
  2. Lateral interactions between CD276 and CD147 are essential for stemness in breast cancer: a novel insight from proximal proteome analysis
    Authors: Seo, YR;Lee, J;Ryu, HS;Kim, EG;Kim, SH;Jeong, J;Jung, H;Jung, Y;Kim, HB;Jo, YH;Kim, YD;Jin, MS;Lee, YY;Kim, KM;Yi, EC;
    Scientific reports
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  3. A clinicopathological analysis of supratentorial ependymoma, ZFTA fusion-positive: utility of immunohistochemical detection of CDKN2A alterations and characteristics of the immune microenvironment
    Authors: Naohito Hashimoto, Tomonari Suzuki, Keisuke Ishizawa, Sumihito Nobusawa, Hideaki Yokoo, Ryo Nishikawa et al.
    Brain Tumor Pathology
  4. CAR T-cell design dependent remodeling of the brain tumor immune microenvironment identify macrophages as key players that inhibit or promote anti-tumor activity
    Authors: Haydar, D;Ibañez-Vega, J;Crawford, J;Chou, CH;Guy, C;Meehl, M;Yi, Z;Langfitt, D;Vogel, P;DeRenzo, C;Gottschalk, S;Roussel, M;Thomas, P;Krenciute, G;
    Research square
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  5. Differential expression of checkpoint markers in the normoxic and hypoxic microenvironment of glioblastomas
    Authors: Stine Asferg Petterson, Mia Dahl Sørensen, Mark Burton, Mads Thomassen, Torben A. Kruse, Signe Regner Michaelsen et al.
    Brain Pathology
  6. Integrative molecular analyses define correlates of high B7-H3 expression in metastatic castrate-resistant prostate cancer
    Authors: Xiaolei Shi, Abderrahman Day, Hannah E. Bergom, Sydney Tape, Sylvan C. Baca, Zoi E. Sychev et al.
    npj Precision Oncology
  7. CRISPR activation screen identifies BCL-2 proteins and B3GNT2 as drivers of cancer resistance to T cell-mediated cytotoxicity
    Authors: J Joung, PC Kirchgatte, A Singh, JH Cho, SP Nety, RC Larson, RK Macrae, R Deasy, YY Tseng, MV Maus, F Zhang
    Nature Communications, 2022-03-25;13(1):1606.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  8. Advanced genetic engineering to achieve in vivo targeting of adenovirus utilizing camelid single domain antibody
    Authors: Myungeun Lee, Zhi Hong Lu, Charles B. Shoemaker, Jacqueline M. Tremblay, Bradley St. St. Croix, Steven Seaman et al.
    Journal of Controlled Release
  9. Evaluation Challenges in the Validation of B7-H3 as Oral Tongue Cancer Prognosticator
    Authors: Meri Sieviläinen, Anna Maria Wirsing, Aini Hyytiäinen, Rabeia Almahmoudi, Priscila Rodrigues, Inger-Heidi Bjerkli et al.
    Head and Neck Pathology
  10. The Inhibition of B7H3 by 2-HG Accumulation Is Associated With Downregulation of VEGFA in IDH Mutated Gliomas
    Authors: Mengli Zhang, Huaichao Zhang, Minjie Fu, Jingwen Zhang, Cheng Zhang, Yingying Lv et al.
    Frontiers in Cell and Developmental Biology
  11. B7-H3 suppresses doxorubicin-induced senescence-like growth arrest in colorectal cancer through the AKT/TM4SF1/SIRT1 pathway
    Authors: R Wang, L Sun, S Xia, H Wu, Y Ma, S Zhan, G Zhang, X Zhang, T Shi, W Chen
    Cell Death & Disease, 2021-05-06;12(5):453.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  12. B7-H3 targeted antibody-based immunotherapy of malignant diseases
    Authors: Theodoros Michelakos, Filippos Kontos, Omar Barakat, Luke Maggs, Joseph H. Schwab, Cristina R. Ferrone et al.
    Expert Opinion on Biological Therapy
  13. CD276 Promotes Vasculogenic Mimicry Formation in Hepatocellular Carcinoma via the PI3K/AKT/MMPs Pathway
    Authors: R Cheng, B Wang, XR Cai, ZS Chen, Q Du, LY Zhou, JM Ye, YL Chen
    Onco Targets Ther, 2020-11-10;13(0):11485-11498.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  14. B7-H3 regulates KIF15-activated ERK1/2 pathway and contributes to radioresistance in colorectal cancer
    Authors: Y Ma, S Zhan, H Lu, R Wang, Y Xu, G Zhang, L Cao, T Shi, X Zhang, W Chen
    Cell Death Dis, 2020-10-03;11(10):824.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  15. Clinical relevance of B7H3 expression in retinoblastoma
    Authors: B Ganesan, S Parameswar, A Sharma, S Krishnakum
    Sci Rep, 2020-06-23;10(1):10185.
    Species: Human
    Sample Types: Tissue Homogenates, Whole Tissue
    Applications: IHC, Western Blot
  16. Evaluation of ductal carcinoma in situ grade via triple-modal molecular imaging of B7-H3 expression
    Authors: Sunitha Bachawal, Gregory R. Bean, Gregor Krings, Katheryne E. Wilson
    npj Breast Cancer
  17. Locoregionally administered B7-H3-targeted CAR T cells for treatment of atypical teratoid/rhabdoid tumors
    Authors: J Theruvath, E Sotillo, CW Mount, CM Graef, A Delaidelli, S Heitzenede, L Labanieh, S Dhingra, A Leruste, RG Majzner, P Xu, S Mueller, DW Yecies, MA Finetti, D Williamson, PD Johann, M Kool, S Pfister, M Hasselblat, MC Frühwald, O Delattre, D Surdez, F Bourdeaut, S Puget, S Zaidi, SS Mitra, S Cheshier, PH Sorensen, M Monje, CL Mackall
    Nat. Med., 2020-04-27;26(5):712-719.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  18. B7-H3 inhibits the IFN-gamma -dependent cytotoxicity of V gamma 9Vδ2 T cells against colon cancer cells
    Authors: Huimin Lu, Tongguo Shi, Mingyuan Wang, Xiaomi Li, Yanzheng Gu, Xueguang Zhang et al.
    OncoImmunology
  19. B7-H3 promotes the cell cycle-mediated chemoresistance of colorectal cancer cells by regulating CDC25A
    Authors: Y Ma, R Wang, H Lu, X Li, G Zhang, F Fu, L Cao, S Zhan, Z Wang, Z Deng, T Shi, X Zhang, W Chen
    J Cancer, 2020-02-03;11(8):2158-2170.
    Species: Human
    Sample Types: Cell Culture Lysates, Whole Tissue
    Applications: IHC, Western Blot
  20. B7-H3 promotes colorectal cancer angiogenesis through activating the NF-kappa B pathway to induce VEGFA expression
    Authors: Ruoqin Wang, Yanchao Ma, Shenghua Zhan, Guangbo Zhang, Lei Cao, Xueguang Zhang et al.
    Cell Death & Disease
  21. Overexpression of B7-H3 in alpha -SMA-Positive Fibroblasts Is Associated With Cancer Progression and Survival in Gastric Adenocarcinomas
    Authors: Shenghua Zhan, Zhiju Liu, Min Zhang, Tianwei Guo, Qiuying Quan, Lili Huang et al.
    Frontiers in Oncology
  22. B7-H3-redirected chimeric antigen receptor T cells target glioblastoma and neurospheres
    Authors: Dean Nehama, Natalia Di Ianni, Silvia Musio, Hongwei Du, Monica Patané, Bianca Pollo et al.
    EBioMedicine
  23. Role of Tumor-Associated Macrophages in the Clinical Course of Pancreatic Neuroendocrine Tumors (PanNETs)
    Authors: Lei Cai, Theodoros Michelakos, Vikram Deshpande, Kshitij S. Arora, Teppei Yamada, David T. Ting et al.
    Clinical Cancer Research
  24. CAR T Cells Targeting B7-H3, a Pan-Cancer Antigen, Demonstrate Potent Preclinical Activity Against Pediatric Solid Tumors and Brain Tumors
    Authors: Robbie G. Majzner, Johanna L. Theruvath, Anandani Nellan, Sabine Heitzeneder, Yongzhi Cui, Christopher W. Mount et al.
    Clinical Cancer Research
  25. B7-H3 promotes aerobic glycolysis and chemoresistance in colorectal cancer cells by regulating HK2
    Authors: 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
    Cell Death Dis, 2019-04-05;10(4):308.
    Species: Human
    Sample Types: Cell Lysates, Whole Tissue
    Applications: IHC-P, Western Blot
  26. B7H3 regulates differentiation and serves as a potential biomarker and theranostic target for human glioblastoma
    Authors: 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
    Lab. Invest., 2019-03-26;0(0):.
    Species: Human
    Sample Types: Cell Lysates, Whole Cells, Whole Tissue
    Applications: ICC, IHC-P, Western Blot
  27. High constitutive B7-H3 expression on human keratinocytes supports T cell immunity
    Authors: D Quandt, E Fiedler, A Müller, WC Marsch, B Seliger
    J. Dermatol. Sci., 2017-04-11;0(0):.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  28. Spectroscopic Photoacoustic Molecular Imaging of Breast Cancer using a B7-H3-targeted ICG Contrast Agent
    Authors: KE Wilson, SV Bachawal, L Abou-Elkac, K Jensen, S Machtaler, L Tian, JK Willmann
    Theranostics, 2017-04-03;7(6):1463-1476.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  29. B7-H3 Promotes the Migration and Invasion of Human Bladder Cancer Cells via the PI3K/Akt/STAT3 Signaling Pathway
    Authors: Y Li, G Guo, J Song, Z Cai, J Yang, Z Chen, Y Wang, Y Huang, Q Gao
    J Cancer, 2017-02-25;8(5):816-824.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P
  30. Immunoregulatory protein B7-H3 reprograms glucose metabolism in cancer cells by ROS-mediated stabilization of HIF-1 alpha
    Authors: Sangbin Lim, Hao Liu, Luciana Madeira Madeira da Silva, Ritu Arora, Zixing Liu, Joshua B. Phillips et al.
    Cancer Research
  31. Preferential Induction of the T Cell Auxiliary Signaling Molecule B7-H3 on Synovial Monocytes in Rheumatoid Arthritis
    Authors: BR Yoon, YH Chung, SJ Yoo, K Kawara, J Kim, IS Yoo, CG Park, SW Kang, WW Lee
    J. Biol. Chem, 2015-12-23;291(8):4048-57.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  32. Expression levels of B7-H3 and TLT-2 in human oral squamous cell carcinoma
    Authors: SHAN-SHAN ZHANG, JING TANG, SHU-YI YU, L I MA, FENG WANG, SHU-LE XIE et al.
    Oncology Letters
  33. Breast Cancer Detection by B7-H3-Targeted Ultrasound Molecular Imaging.
    Authors: Bachawal S, Jensen K, Wilson K, Tian L, Lutz A, Willmann J
    Cancer Res, 2015-04-21;75(12):2501-9.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P
  34. Origination of new immunological functions in the costimulatory molecule B7-H3: the role of exon duplication in evolution of the immune system.
    Authors: Sun J, Fu F, Gu W, Yan R, Zhang G, Shen Z, Zhou Y, Wang H, Shen B, Zhang X
    PLoS ONE, 2011-09-13;6(9):e24751.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  35. Anterior pituitary progenitor cells express costimulatory molecule 4Ig-B7-H3.
    Authors: Nagai Y, Aso H, Ogasawara H, Tanaka S, Taketa Y, Watanabe K, Ohwada S, Rose MT, Kitazawa H, Yamaguchi T
    J. Immunol., 2008-11-01;181(9):6073-81.
    Species: Bovine
    Sample Types: Cell Lysates
    Applications: Western Blot
  36. Interactions of T cells with fibroblast-like synoviocytes: role of the B7 family costimulatory ligand B7-H3.
    Authors: Tran CN, Thacker SG, Louie DM, Oliver J, White PT, Endres JL, Urquhart AG, Chung KC, Fox DA
    J. Immunol., 2008-03-01;180(5):2989-98.
    Species: Human
    Sample Types: Whole Cells, Whole Tissue
    Applications: ICC, IHC-Fr, IHC-P
  37. Tumor cell and tumor vasculature expression of B7-H3 predict survival in clear cell renal cell carcinoma.
    Authors: Crispen PL, Sheinin Y, Roth TJ, Lohse CM, Kuntz SM, Frigola X, Thompson RH, Boorjian SA, Dong H, Leibovich BC, Blute ML, Kwon ED
    Clin. Cancer Res., 2008;14(16):5150-7.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC Paraffin-embedded
  38. 12-O-tetradecanoyl phorbol 13-acetate induces the expression of B7-DC, -H1, -H2, and -H3 in K562 cells.
    Authors: Jang BC, Park YK, Choi IH, Kim SP, Hwang JB, Baek WK, Suh MH, Mun KC, Suh SI
    Int. J. Oncol., 2007-12-01;31(6):1439-47.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  39. The immunomodulatory proteins B7-DC, B7-H2, and B7-H3 are differentially expressed across gestation in the human placenta.
    Authors: Petroff MG, Kharatyan E, Torry DS, Holets L
    Am. J. Pathol., 2005-08-01;167(2):465-73.
    Species: Human
    Sample Types: Tissue Homogenates, Whole Tissue
    Applications: IHC-P, Western Blot
  40. Assessment of combined expression of B7-H3 and B7-H4 as prognostic marker in esophageal cancer patients.
    Authors: Chen L, Xie Q, Wang Z et al.
    Oncotarget.

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Human B7-H3 Antibody
By Anonymous on 11/02/2018
Application: Simple Western Sample Tested: b7h3 4ig protein,Purified protein Species: Human
Simple Western Human B7-H3 Antibody AF1027