Human B7-H3 MAb (Clone 185504)

Catalog # Availability Size / Price Qty
MAB1027-SP
MAB1027-100
MAB1027-500
Detection of Human B7‑H3 by Western Blot.
4 Images
Product Details
Citations (6)
FAQs
Supplemental Products
Reviews (4)

Human B7-H3 MAb (Clone 185504) Summary

Species Reactivity
Human
Specificity
Detects human B7-H3 in direct ELISAs and Western blots. In Western blots, no cross-reactivity with recombinant human (rh) B7‑H1, rhB7‑H2, rhB7‑1, rhB7‑2, or recombinant mouse PD-L2 is observed.
Source
Monoclonal Mouse IgG1 Clone # 185504
Purification
Protein A or G purified from hybridoma culture supernatant
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
2 µg/mL
See below
Flow Cytometry
0.25 µg/106 cells
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 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 2 µg/mL of Mouse Anti-Human B7-H3 Monoclonal Antibody (Catalog # MAB1027) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). 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 cancer cell line was stained with Mouse Anti-Human B7-H3 Monoclonal Antibody (Catalog # MAB1027, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')2Secondary Antibody (Catalog # F0102B).

Flow Cytometry Detection of B7-H3 antibody in Human Dendritic Cells antibody by Flow Cytometry. View Larger

Detection of B7‑H3 in Human Dendritic Cells by Flow Cytometry. Human monocyte-derived dendritic cells were stained with Mouse Anti-Human B7-H3 Monoclonal Antibody (Catalog # MAB1027, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B).

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 2 µg/mL of Mouse Anti-Human B7-H3 Monoclonal Antibody (Catalog # MAB1027) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). 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 # MAB5718) 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 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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); 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

Citations for Human B7-H3 MAb (Clone 185504)

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.

6 Citations: Showing 1 - 6
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  1. Targeting B7-H3 via chimeric antigen receptor T cells and bispecific killer cell engagers augments antitumor response of cytotoxic lymphocytes
    Authors: J Liu, S Yang, B Cao, G Zhou, F Zhang, Y Wang, R Wang, L Zhu, Y Meng, C Hu, H Liang, X Lin, K Zhu, G Chen, KQ Luo, L Di, Q Zhao
    Journal of hematology & oncology, 2021;14(1):21.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  2. Targeting B7-H3 Immune Checkpoint With Chimeric Antigen Receptor-Engineered Natural Killer Cells Exhibits Potent Cytotoxicity Against Non-Small Cell Lung Cancer
    Authors: S Yang, B Cao, G Zhou, L Zhu, L Wang, L Zhang, HF Kwok, Z Zhang, Q Zhao
    Front Pharmacol, 2020;11(0):1089.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Immunoprecipitation
  3. Genetic evolution of T-cell resistance in the course of melanoma progression.
    Authors: Sucker A, Zhao F, Real B, Heeke C, Bielefeld N, Mabetaen S, Horn S, Moll I, Maltaner R, Horn P, Schilling B, Sabbatino F, Lennerz V, Kloor M, Ferrone S, Schadendorf D, Falk C, Griewank K, Paschen A
    Clin Cancer Res, 2014;20(24):6593-604.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  4. Multiple inhibitory ligands induce impaired T-cell immunologic synapse function in chronic lymphocytic leukemia that can be blocked with lenalidomide: establishing a reversible immune evasion mechanism in human cancer.
    Authors: Ramsay, Alan G, Clear, Andrew J, Fatah, Rewas, Gribben, John G
    Blood, 2012;120(7):1412-21.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  5. 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: Flow Cytometry
  6. Expression of B7-H1 on gastric epithelial cells: its potential role in regulating T cells during Helicobacter pylori infection.
    Authors: Das S, Suarez G, Beswick EJ, Sierra JC, Graham DY, Reyes VE
    J. Immunol., 2006;176(5):3000-9.
    Species: Human
    Sample Types: Cell Culture Supernates
    Applications: Western Blot

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Reviews for Human B7-H3 MAb (Clone 185504)

Average Rating: 4.5 (Based on 4 Reviews)

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Human B7-H3 MAb (Clone 185504)
By Anonymous on 06/22/2022
Application: Western Blot Sample Tested: Osteosarcoma U2OS cell line Species: Human

Human B7-H3 Antibody
By Anonymous on 05/01/2020
Application: Simple Western Sample Tested: Purified protein Species: Human

Human B7-H3 Antibody
By Jenna Nguyen on 05/07/2019
Application: Functional Assay Sample Tested: 3T3-L1 mouse embryonic fibroblast adipose-like cell line Species: Mouse

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