Human B7-H3 Antibody (185504)

R&D Systems | Catalog # MAB1027

R&D Systems
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Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Knockout Validated, Western Blot, Flow Cytometry, CyTOF-ready

Cited:

Immunohistochemistry, Western Blot, Flow Cytometry, Immunoprecipitation

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 185504
View all B7-H3 Primary Antibodies »

Product Specifications

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 Western blots, no cross-reactivity with recombinant human (rh) B7‑H1, rhB7‑H2, rhB7‑1, rhB7‑2, or recombinant mouse PD-L2 is observed.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Human B7-H3 Antibody (185504)

Detection of Human B7-H3 antibody by Western Blot.

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

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

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).
Western Blot Shows Human B7-H3 Antibody Specificity by Using Knockout Cell Line.

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.
Detection of B7-H3 by Flow Cytometry

Detection of B7-H3 by Flow Cytometry

Functional analysis of B7-H3 CAR-T cells. a Cytokine production in B7-H3 CAR and vehicle T cells. Effector T cells were incubated with target A549 or HCT 116 cells at a ratio of 10:1 for 24 h. The concentrations of cytokines were measured using flow cytometry. b Memory cell phenotypes of B7-H3 CAR-T cells. Effector T cells were incubated with target A549 or HCT 116 cells at a ratio of 10:1 for 24 h. Expression of CD62L and CCR7 on T cells was identified with staining with anti-CD62L and anti-CCR7 antibodies, respectively, and measured using flow cytometry. c Degranulation markers in B7-H3 CAR T cells. Effector T cells were incubated with target A549 or HCT 116 cells at a ratio of 10:1 for 6 h (CD107a) or 24 h (granzyme B and perforin). Expressions of CD107a, intracellular granzyme B and perforin staining on T cells were stained with anti CD107a, granzyme B and perforin antibodies and measured by flow cytometry. Data from 2–3 independent experiments are presented as the mean ± standard deviation. *p < 0.05, **p < 0.01, ***p < 0.001. The p values were analyzed using a Student’ t test Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33514401), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human B7-H3 Antibody (185504)

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/106 cells
Sample: Human monocyte-derived dendritic cells and PC‑3 human prostate cancer cell line

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.

Western Blot

2 µg/mL
Sample: LNCaP human prostate cancer cell line and U2OS human osteosarcoma cell line

Reviewed Applications

Read 4 reviews rated 4.5 using MAB1027 in the following applications:

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Advanced Features

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  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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.

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.

Calculators

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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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

Alternate Names

B7H3, CD276

Entrez Gene IDs

80381 (Human); 102657 (Mouse); 315716 (Rat); 102131295 (Cynomolgus Monkey)

Gene Symbol

CD276

UniProt

NP_079516

Additional B7-H3 Products

Product Documents for Human B7-H3 Antibody (185504)

Certificate of Analysis

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Product Specific Notices for Human B7-H3 Antibody (185504)

For research use only

Citations for Human B7-H3 Antibody (185504)

Customer Reviews for Human B7-H3 Antibody (185504) (4)

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Customer Images

Showing  1 - 4 of 4 reviews Showing All
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  • Human B7-H3 MAb (Clone 185504)
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Osteosarcoma U2OS cell line
    Species: Human
    Verified Customer | Posted 06/22/2022
    Human B7-H3 Antibody (185504) MAB1027
  • Human B7-H3 Antibody
    Name: Anonymous
    Application: Simple Western
    Sample Tested: Purified protein
    Species: Human
    Verified Customer | Posted 05/01/2020
    Human B7-H3 Antibody (185504) MAB1027
  • Human B7-H3 Antibody
    Name: Jenna Nguyen
    Application: Functional Assay
    Sample Tested: 3T3-L1 mouse embryonic fibroblast adipose-like cell line
    Species: Mouse
    Verified Customer | Posted 05/07/2019
  • Human B7-H3 Antibody
    Name: Anonymous
    Application: Simple Western
    Sample Tested: Purified protein
    Species: Human
    Verified Customer | Posted 11/02/2018
    Human B7-H3 Antibody (185504) MAB1027

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