HeLa human cervical epithelial carcinoma cell line was stained with Mouse Anti-Human B7-H6 Monoclonal Antibody (Catalog # MAB71442, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). View our protocol for Staining Membrane-associated Proteins.

Detection of B7‑H6 in HEK293 Human Cell Line Transfected with Human B7-H6 and eGFP by Flow Cytometry.

HEK293 human embryonic kidney cell line transfected with either (A) human B7-H6 or (B) irrelevant transfectants and eGFP were stained with Mouse Anti-Human B7-H6 Monoclonal Antibody (Catalog # MAB71442) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). View our protocol for Staining Membrane-associated Proteins.

Human B7‑H6 ELISA Standard Curve.

Recombinant Human B7-H6 protein was serially diluted 2-fold and captured by Mouse Anti-Human B7-H6 Monoclonal Antibody (Catalog # MAB7144) coated on a Clear Polystyrene Microplate (Catalog # DY990). Mouse Anti-Human B7-H6 Monoclonal Antibody (Catalog # MAB71442) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994).

Applications for Human B7-H6 Antibody

Application

Recommended Usage

CyTOF-ready

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

ELISA

This antibody functions as an ELISA detection antibody when paired with Mouse Anti-Human B7‑H6 Monoclonal Antibody (Catalog # MAB7144).

This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human B7-H6 DuoSet ELISA (Catalog # DY7144-05) for convenient development of a sandwich ELISA.

Flow Cytometry

0.25 µg/10⁶ cells
Sample: HeLa human cervical epithelial carcinoma cell line and HEK293 Human Cell Line Transfected with Human B7-H6 and eGFP

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

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-H6

B7-H6 is a glycosylated member of the B7 family of immune co‑stimulatory proteins (1, 2). Mature human B7-H6 consists of a 238 amino acid (aa) extracellular domain (ECD) that contains one Ig-like V domain and one Ig-like C1 domain, a 21 aa transmembrane segment, and a 171 aa cytoplasmic domain that contains one ITIM, one SH2, and one SH3 motif (3). Both of the Ig-like domains carry N-linked glycosylation (4). Within the ECD, human B7-H6 shares 99%, 94%, and 87% aa sequence identity with chimpanzee, orangutan, and gibbon B7-H6, respectively, and 53% - 56% with bovine, canine, and equine B7-H6. Orthologs in mouse and rat have not been identified. The Ig-like V domain mediates 1:1 stoichiometric binding of B7-H6 to NKp30 expressed on NK cells (4, 5). It does not show binding to NKp44, NKp46, or NKG2D (3, 6). Ligation of NKp30 by B7-H6 induces NK cell activation and target cell cytolysis (3). B7-H6 is expressed on a wide range of hematopoietic, carcinoma, and melanoma tumor cells, which is consistent with the detection of NKp30 binding sites on many tumors (3, 7). The expression of NKp30 ligands on tumor cells correlates with tumor cell sensitivity to NKp30‑dependent cell lysis (7).

References

Zou, W. and L. Chen (2008) Nat. Rev. Immunol. 8:467.
Bour-Jordan, H. et al. (2011) Immunol. Rev. 241:180.
Brandt, C.S. et al. (2009) J. Exp. Med. 206:1495.
Li, Y. et al. (2011) J. Exp. Med. 208:703.
Joyce, M.G. et al. (2011) Proc. Natl. Acad. Sci. 108:6223.
Arnon, T.I. et al. (2006) Semin. Cancer Biol. 16:348.
Byrd, A. et al. (2007) PLoS ONE 2:e1339.

Long Name

B7 Homolog 6

Alternate Names

B7H6, DKFZp686I21167, DKFZp686O24166, NCR3LG1

Entrez Gene IDs

374383 (Human); 102121659 (Cynomolgus Monkey)

Gene Symbol

NCR3LG1

UniProt

Q68D85

Additional B7-H6 Products

Product Documents for Human B7-H6 Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human B7-H6 Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human B7-H6 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB71442

Key Product Details

Species Reactivity

Validated:

Applications

Label

Antibody Source

View all B7-H6 Antibodies »

Product Summary for Human B7-H6 Antibody

Immunogen

Specificity

Clonality

Host

Isotype

Scientific Data Images for Human B7-H6 Antibody

Detection of B7‑H6 in HeLa Human Cell Line by Flow Cytometry.