Human B7-H4 Antibody

Recombinant Monoclonal Antibody
Catalog # Availability Size / Price Qty
MAB65761R-100
MAB65761R-SP
Human B7-H4 Antibody in Flow Cytometry
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Human B7-H4 Antibody Summary

Species Reactivity
Human
Specificity
Detects human B7-H4 in direct ELISAs.
Source
Recombinant Monoclonal Mouse IgG2A Clone # 973816R
Purification
Protein A or G purified from cell culture supernatant
Immunogen
Mouse myeloma cell line NS0-derived recombinant human B7-H4
Phe29-Ala258
Accession # Q7Z7D3
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.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Flow Cytometry
0.25 µg/106 cells
See below
Immunohistochemistry
1-25 µg/mL
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.

Data Examples

Flow Cytometry Detection of B7-H4 in HEK293 Human Cell Line Transfected with Human B7-H4 and eGFP by Flow Cytometry. View Larger

Detection of B7-H4 in HEK293 Human Cell Line Transfected with Human B7-H4 and eGFP by Flow Cytometry. HEK293 human embryonic kidney cell line transfected with either (A) human B7-H4 or (B) irrelevant transfectants and eGFP were stained with Mouse Anti-Human B7‑H4 Monoclonal Antibody (Catalog # MAB65761R) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B).View our protocol for Staining Membrane-associated Proteins.

Immunohistochemistry B7-H4  in Human Breast Cancer Tissue. View Larger

B7-H4 in Human Breast Cancer Tissue. B7‑H4 was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Mouse Anti-Human B7‑H4 Monoclonal Antibody (Catalog # MAB65761R) at 1.7 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

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.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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-H4

B7-H4, also known as VTCN1, B7x and B7S1, is a 50‑80 kDa glycosylated member of the BTN/MOG family of immunomodulatory protein (1, 2). Mature human B7-H4 consists of a 235 amino acid (aa) extracellular domain (ECD) with one Ig-like V-set domain and one Ig-like C2-set domain, a 21 aa transmembrane segment, and a 2 aa cytoplasmic tail (3-5). Within the ECD, human B7-H4 shares 90% aa sequence identity with mouse and rat B7-H4. It shares 22%-28% aa sequence identity with human B7-1, B7-2, B7-H1, B7-H2, B7-H3, and PD‑L2. Alternate splicing of human B7-H4 generates an additional isoform that lacks the first Ig-like domain. B7-H4 is expressed on the surface of activated lymphocytes, macrophages, monocytes, dendritic cells, epithelial cells, and bone marrow-derived mesenchymal stem cells (4-8). Following binding to activated T cells, B7-H4 serves as a co‑inhibitor of the T cell response. This is accomplished by reverse signaling that can induce either cell cycle arrest, or apoptosis in B7-H4 expressing cells (3-5, 9, 10). B7‑H4 is up‑regulated in several carcinomas in correlation with tumor progression and metastasis (2, 7, 11, 12). A soluble form of B7-H4 is elevated in the serum of ovarian cancer, renal cell carcinoma, and rheumatoid arthritis patients, also in correlation with advanced disease status (13-15). Soluble B7‑H4 functions as a decoy molecule that blocks the inhibitory influence of B7‑H4 on immune activation (15). Despite evidence for the involvement of B7-H4 in immune regulation, mice deficient in its expression do not show significant immune deficiencies, suggesting compensation by other molecules in vivo (16).

References
  1. Yi, K.H. and L. Chen (2009) Immunol. Rev. 229:145.
  2. Salceda, S. et al. (2005) Exp. Cell Res. 306:128.
  3. Zang, X. et al. (2003) Proc. Natl. Acad. Sci. 100:10388.
  4. Prasad, V.R. et al. (2003) Immunity 18:863.
  5. Sica, G.L. et al. (2003) Immunity 18:849.
  6. Kryczek, I. et al. (2006) J. Exp. Med. 203:871.
  7. Tringler, B. et al. (2005) Clin. Cancer Res. 11:1842.
  8. Xue, Q. et al. (2010) Stem Cells Dev. 19:27.
  9. Song, H. et al. (2008) Cancer Lett. 266:227.
  10. Park, G.B. et al. (2009) Immunology 128:360.
  11. Zang, X. et al. (2007) Proc. Natl. Acad. Sci. 104:19458.
  12. Krambeck, A.E. et al. (2006) Proc. Natl. Acad. Sci. 103:10391.
  13. Simon, I. et al. (2006) Cancer Res. 66:1570.
  14. Thompson, R.H. et al. (2008) Cancer Res. 68:6054.
  15. Azuma, T. et al. (2009) PloS Med. 6:e1000166.
  16. Suh, W.-K. et al. (2006) Mol. Cell. Biol. 26:6403.
Long Name
B7 Homolog 4
Entrez Gene IDs
79679 (Human); 242122 (Mouse); 295322 (Rat)
Alternate Names
B7h.5; B7H4; B7-H4; B7H4T-cell costimulatory molecule B7x; B7S1; B7S1VCTN1; B7x; B7XPRO1291; FLJ22418; Immune costimulatory protein B7-H4; Protein B7S1; T cell costimulatory molecule B7x; V-set domain containing T cell activation inhibitor 1; V-set domain-containing T-cell activation inhibitor 1; Vtcn1

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FAQs

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Cell and Tissue Staining Kits

Immunohistochemistry Reagents

Isotype Controls

Mouse IgG2A Isotype Control

Reconstitution Buffers

Secondary Antibodies

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