Human B7-H6 Antibody

Catalog # Availability Size / Price Qty
MAB71442-100
MAB71442-SP
Detection of B7‑H6 in HeLa Human Cell Line by Flow Cytometry.
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Human B7-H6 Antibody Summary

Species Reactivity
Human
Specificity
Detects human B7-H6 in direct ELISAs.
Source
Monoclonal Mouse IgG1 Clone # 875002
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Mouse myeloma cell line NS0-derived recombinant human B7-H6
Met1-Ser262
Accession # Q68D85
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
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
See below
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 

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.

Scientific Data

Flow Cytometry Detection of B7-H6 antibody in HeLa Human Cell Line antibody by Flow Cytometry. View Larger

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

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

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).

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.
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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
  1. Zou, W. and L. Chen (2008) Nat. Rev. Immunol. 8:467.
  2. Bour-Jordan, H. et al. (2011) Immunol. Rev. 241:180.
  3. Brandt, C.S. et al. (2009) J. Exp. Med. 206:1495.
  4. Li, Y. et al. (2011) J. Exp. Med. 208:703.
  5. Joyce, M.G. et al. (2011) Proc. Natl. Acad. Sci. 108:6223.
  6. Arnon, T.I. et al. (2006) Semin. Cancer Biol. 16:348.
  7. Byrd, A. et al. (2007) PLoS ONE 2:e1339.
Long Name
B7 Homolog 6
Entrez Gene IDs
374383 (Human); 102121659 (Cynomolgus Monkey)
Alternate Names
B7 homolog 6; B7H6; B7-H6; DKFZp686I21167; DKFZp686O24166; NCR3LG1

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