Human B7-H7/HHLA2 Antibody

MAB80841 Datasheet / COA / SDS

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MAB80841
MAB80841-SP

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Detection of B7-H7/HHLA2 in HEK Human Cell Line Transfected with Human B7-H7/HHLA2 and eGFP by Flow Cytometry.

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Human B7-H7/HHLA2 Antibody Summary

Species Reactivity

Human

Specificity

Detects human B7-H7/HHLA2 in direct ELISAs.

Source

Monoclonal Mouse IgG₁ Clone # 907812

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

Human embryonic kidney cell line HEK293-derived human B7-H7/HHLA2
Met1-Asn344
Accession # Q9UM44

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Flow Cytometry

0.25 µg/10⁶ cells

See below

Blockade of Receptor-ligand Interaction

In a functional ELISA, 100-300 ng/mL of Mouse anti-Human B7-H7/HHLA2 Antibody (Catalog # MAB80841) will block 50% of the binding of 500 ng/mL of Recombinant Human B7-H7/HHLA2 Fc Chimera (Catalog # 8084-B7) to immobilized Recombinant Human TMIGD2/CD28H Fc Chimera (Catalog # 8316-TR) coated at 125 ng/mL
(100 µL/well). At 1.25 μg/mL, this antibody will block >90% of the binding.

CyTOF-ready

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

Immunocytochemistry

8-25 µg/mL

See below

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.

Data Examples

Flow Cytometry

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Detection of B7-H7/HHLA2 in HEK Human Cell Line Transfected with Human B7-H7/HHLA2 and eGFP by Flow Cytometry. HEK293 human embryonic kidney cell line transfected with either (A) human B7-H7/HHLA2 or (B) irrelevant protein, and eGFP was stained with Mouse Anti-Human B7-H7/HHLA2 Monoclonal Antibody (Catalog # MAB80841) followed by anti-Mouse IgG APC-conjugated secondary antibody (Catalog # F0101B). Quadrant markers were set based on control antibody staining (Catalog # MAB002). View our protocol for Staining Membrane-associated Proteins.

Immunocytochemistry

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B7-H7/HHLA2 in Human PBMCs.
B7-H7/HHLA2 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Mouse Anti-Human B7-H7/HHLA2 Monoclonal Antibody (Catalog # MAB80841) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Blockade of Receptor-ligand Interaction

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TMIGD2/CD28H Binding to B7-H7/
HHLA2-transfected HEK293 Human Cell Line is Blocked by Human B7-H7/HHLA2 Antibody. In a functional flow cytometry test, biotinylated recombinant human TMIGD2/
CD28H (10 ng/mL, Catalog # 8316-TR) binds to HEK293 human embryonic kidney cell line transfected with human B7-H7/
HHLA2 (black dotted line). Binding is completely blocked (orange histogram) by 2.5 μg/mL of Mouse Anti-Human B7-H7/
HHLA2 Monoclonal Antibody (Catalog # MAB80841). Mouse IgG1 Isotype Control (Catalog # MAB002) at 2.5 μg/mL was used as a control (blue line). Cells were stained with Streptavidin-APC (Catalog # F0050).

Blockade of Receptor-ligand Interaction

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B7-H7 Binding to TMIGD2/
CD28H Blocked by Human
B7-H7/HHLA2 Antibody. In a Surface Plasmon Resonance test, Recombinant Human TMIGD2/
CD28H Fc Chimera (Catalog # 8316-TR), coated at 10 ng/mL binds to Recombinant Human
B7-H7/HHLA2 Fc Chimera (Catalog # 8084-B7, 5 μg/mL) (blue line). Binding is completely blocked (red) by 50 μg/mL of Mouse Anti-Human B7-H7/
HHLA2 Monoclonal Antibody (Catalog # MAB80841). Mouse IgG1 isotype (Catalog # MAB002) and irrelevant mouse monoclonal antibody were used as controls and did not affect TMIGD2/
CD28H binding to B7-H7/HHLA2 (not shown).

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Reconstitution Buffer Available

Reconstitution Buffer 1 (PBS)

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C

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-H7/HHLA2

B7-H7, also known as HHLA2 (HERV-H LTR-associating 2), is a member of the B7 family of immune regulatory proteins (1, 2). Mature human B7-H7 consists of a 322 amino acid (aa) extracellular domain (ECD) with three immunoglobulin-like domains, a 21 aa transmembrane segment, and a 49 aa cytoplasmic domain (3-5).
B7‑H7 is constitutively expressed on monocytes and is up-regulated by LPS and IFN-gamma stimulation. It is expressed on LPS/IFN-gamma treated B cells but not on resting B cells (5). B7-H7 binds to cell surface determinants on resting and mature T cells, B cells, and monocytes as well as on immature and mature dendritic cells (5). Soluble B7-H7 inhibits the proliferation of activated CD4⁺ and CD8⁺ T cells and their production of IFN-gamma, TNF-alpha, IL-5, IL-10, IL-13, IL-17A, and IL-22 (5).

References

Zou, W. and L. Chen (2008) Nat. Rev. Immunol. 8:467.
Bour-Jordan, H. et al. (2011) Immunol. Rev. 241:180.
Mager, D.L. et al. (1999) Genomics 59:255.
Flajnik, M.M. et al. (2012) Immunogenetics 64:571.
Zhao, R. et al. (2013) Proc. Natl. Acad. Sci. USA 110:9879.

Long Name

B7 Homolog 7

Entrez Gene IDs

11148 (Human); 102137704 (Cynomolgus Monkey)

Alternate Names

B7H7; B7-H7; HERV-H LTR-associating 2; HERV-H LTR-associating protein 2; HHLA2; Human endogenous retrovirus-H long terminal repeat-associating protein 2