Human B7-H3 Antibody

(10 citations)   
  • Species Reactivity
    Human
  • 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.
  • 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 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.
Data Examples
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.
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 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.
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 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 90-110 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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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); 102131295 (Cynomolgus Monkey)
  • Alternate Names:
    B7H3; B7-H3; B7H34Ig-B7-H3; B7-H3B7 homolog 3; CD276 antigen; CD276 molecule; CD276; Costimulatory molecule
Related Research Areas
Citations:

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.

10 Citations: Showing 1 - 10
Filter your results:

Species
Applications
Sample Type
  1. 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;8(5):816-824.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  2. 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;7(6):1463-1476.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC
  3. 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, 2016;291(8):4048-57.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  4. 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;75(12):2501-9.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  5. 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;6(9):e24751.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  6. 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 Type: Whole Tissue
    Application: IHC Paraffin-embedded
  7. 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;181(9):6073-81.
    Species: Bovine
    Sample Type: Cell Lysates
    Application: WB
  8. 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;180(5):2989-98.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  9. 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;31(6):1439-47.
    Species: Human
    Sample Type: Whole Cells
    Application: Flow
  10. 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;167(2):465-73.
    Species: Human
    Sample Type: Tissue Homogenates
    Application: WB
Cell and Tissue Staining Kits
Description Application Cat# Citations Images  

Anti-Goat HRP-DAB Cell & Tissue Staining Kit

CTS008 13
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....
Immunohistochemistry Reagents
Description Application Cat# Citations Images  

Antigen Retrieval Reagent-Universal

CTS015 10
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Antigen Retrieval Reagent-Basic

CTS013 4
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Mounting Medium

CTS011 4
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Antigen Retrieval Reagent Sampler (50 mL each)

CTS016
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Antigen Retrieval Reagent-Acidic

CTS014
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

NorthernLights Guard Mounting Media

NL996
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

VisUCyte Antigen Retrieval Reagent-Basic

VCTS021
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

VisUCyte Antigen Retrieval Reagent-Acidic

VCTS022
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

VisUCyte Antigen Retrieval Reagent-Universal

VCTS023
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....
Isotype Controls
Description Application Cat# Citations Images  

Normal Goat IgG Control

Ctrl AB-108-C 191  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....
Secondary Antibodies
Description Application Cat# Citations Images  

Goat IgG HRP-conjugated Antibody

WB, Simple Western HAF109 19  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Goat IgG HRP-conjugated Antibody

WB HAF017 15  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Goat IgG (H+L) PE-conjugated Antibody

Flow F0107 4  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Donkey Anti-Goat IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL001 9
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Donkey Anti-Goat IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL003 5
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Goat IgG (H+L) APC-conjugated Antibody

Flow F0108 6  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Goat IgG Horseradish Peroxidase-conjugated Antibody

WB HAF019 6  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Donkey Anti-Goat IgG Antibody

WB AF109 6
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Donkey Anti-Goat IgG Biotinylated Antibody

WB BAF109 3
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Goat IgG VisUCyte HRP Polymer

IHC VC004  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Goat IgG (H+L) Fluorescein-conjugated Antibody

Flow F0109 2  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Donkey Anti-Goat IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL002
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Chicken Anti-Goat IgG Biotinylated Antibody

WB BAF019
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Rabbit Anti-Goat IgG Biotinylated Antibody

WB BAF017
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Rabbit Anti-Goat IgG (H+L) Affinity Purified PAb, X Absorbed

WB, ELISA R-401-C-ABS
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Donkey Anti-Goat IgG (H+L) PerCP-conjugated Antibody

Flow F0124
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Have you used Human B7-H3 Antibody?

Submit a review and receive a $25US/€18/£15/$25CAN amazon gift card if you include an image - $10US/€7/£6/$10CAN Amazon card for reviews without an image. Limited to verified customers in USA, Canada and Europe.

Order Details

Contact Information

Calculators

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.

=
÷
R&D Systems Guarantee

Monoclonal
Antibody
Panels For
Screening

Learn More
Monoclonal Antibodies

Feedback

Customer Information