Human B7-H3 Antibody

Name: Human B7-H3 Antibody
Brand: R&D Systems
SKU: AF1027-SP
Rating: 4 (1 reviews)

AF1027 12 Citations 1 Review Datasheet / COA / SDS

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AF1027
AF1027-SP

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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. 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

Western Blot

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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.

Immunohistochemistry

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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.

Flow Cytometry

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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).

Simple Western

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Detection of Human B7‑H3 by Simple Western^TM. 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.

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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

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Reconstitution Buffer Available

Reconstitution Buffer 1 (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

Chapoval, A.I. et al. (2001) Nat. Immunol. 2:269.
Sharpe, A.H. and G.J. Freeman (2002) Nat. Rev. Immunol. 2:116.
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

Product Datasheets

Product Datasheet

COA

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Related Research Areas

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.

12 Citations: Showing 1 - 10
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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;0(0):.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot
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;10(4):308.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot
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;0(0):.
Species: Human
Sample Types: Whole Tissue
Applications: IHC
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 Types: Whole Tissue
Applications: IHC-P
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 Types: Whole Tissue
Applications: IHC
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 Types: Cell Lysates
Applications: Western Blot
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 Types: Whole Tissue
Applications: IHC-P
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 Types: Cell Lysates
Applications: Western Blot
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 Types: Whole Cells
Applications: ICC
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 Types: Cell Lysates
Applications: Western Blot

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 Types: Whole Cells
Applications: Flow Cytometry

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 Types: Tissue Homogenates
Applications: Western Blot