Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Knockout Validated, ELISA, Flow Cytometry, CyTOF-ready

Cited:

Flow Cytometry

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 875001
View all B7-H6 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human B7-H6
Met1-Ser262
Accession # Q68D85

Specificity

Detects human B7-H6 in ELISA.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Human B7‑H6 Antibody

Human B7-H6 Antibody in ELISA Standard Curve.

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 (DY990). Mouse Anti-Human B7-H6 Monoclonal Antibody (MAB71442) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (DY994).
B7-H6 Antibody Specificity is Shown by Flow Cytometry in Knockout Cell Line.

B7-H6 Specificity is Shown by Flow Cytometry in Knockout Cell Line.

B7-H6 knockout HeLa human cervical epithelial carcinoma cell line was stained with Mouse Anti-Human B7-H6 Monoclonal Antibody (Catalog # MAB7144, filled histogram) or isotype control antibody (MAB002, open histogram) followed by anti-Mouse IgG PE-conjugated secondary antibody (F0102B). No staining in the B7-H6 knockout HeLa cell line was observed. View our protocol for Staining Membrane-associated Proteins.

Detection of B7‑H6 in K562 cells by Flow Cytometry.

K562 cells were stained with Mouse Anti-Human B7‑H6 Monoclonal Antibody (Catalog # MAB7144, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). View our protocol for Staining Membrane-associated Proteins.
Detection of Human B7-H6 by Flow Cytometry

Detection of Human B7-H6 by Flow Cytometry

Validation of cell surface proteins regulated by the US12 family.Flow cytometry confirmed proteomic data for proteins representative of each category enriched in the DAVID analysis. Staining in mock/US12 family member deletion mutant infections (blue line) is shown relative to the parental HCMV infection (red line). Flow cytometry was carried out for cell surface expression of MHC I (W6/32) as a control for HCMC infection and isotype antibody staining controls (with directly PE-conjugated IgG1, IgG2a or IgG2b antibodies or for unconjugated antibodies mIgG and an anti-mouse-AF647 conjugated secondary antibody). Infected cells were assessed by the % cells with down-regulated MHC I compared to the mock-infected cells (HCMV 94%, delta US12 94%, delta US13 94%, delta US14 83%, delta US15 95%, delta US16 97%, delta US17 85%, delta US18 94%, delta US19 96%, delta US20 94%, delta US21 94%, delta US12-21 96%). Results are representative of at least two independent experiments.DOI:https://dx.doi.org/10.7554/eLife.22206.013 Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28186488), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 capture antibody when paired with Mouse Anti-Human B7‑H6 Monoclonal Antibody (Catalog # MAB71442).

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: K562 human chronic myelogenous leukemia cell line

Knockout Validated

B7-H6 is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in B7-H6 knockout HeLa cell line.

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • 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-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

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

Certificate of Analysis

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Product Specific Notices for Human B7‑H6 Antibody

For research use only

Citations for Human B7‑H6 Antibody

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Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for Human B7‑H6 Antibody

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  • Q: I would like to use this antibody on Western Blot. Do you have any experience with this antibody in this technique? I see that only Flow Cytometry and ELISA are described in the datasheet.

    A:

    FYI, this antibody was produced by our sister company, R & D System. Unfortunately MAB7144 has never been tested in WB (only be validated in flow and ELISA) and therefore we don't know whether this antibody can work in the WB experiments; no protocol could be suggested to you, apologize for any inconvenience. Although we have another anti-human B7-H6 antibody, but it has only been validated in IHC-P (not WB). If you are willing to purchase and use either antibody in WB, you will be automatically qualified for our Innovator's Reward Program, which will award you the next antibody free of charge to you.

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