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 MAb (Clone 185504)

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 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 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).

View 1 more images

Applications for Human B7-H3 MAb (Clone 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

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

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

Functional Assay (1 Review)
Simple Western (2 Reviews)
Western Blot (1 Review)

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.

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

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 MAb (Clone 185504)

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human B7-H3 MAb (Clone 185504)

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human B7-H3 MAb (Clone 185504)

R&D Systems, part of Bio-Techne | Catalog # MAB1027

Key Product Details

Validated by

Species Reactivity

Validated:

Cited:

Applications