B7-H4 Antibody (2318A) [Unconjugated]

HEK293 human embryonic kidney cell line transfected with human B7-H4 and eGFP was stained with either (A) Rabbit Anti-Human B7-H4 Monoclonal Antibody (Catalog # MAB65764) or (B) Rabbit IgG control antibody (Catalog # MAB1050) followed by APC-conjugated Goat anti-Rabbit IgG Secondary Antibody (Catalog # F0111). View our protocol for Staining Membrane-associated Proteins.

Western blot shows lysates of SK‑BR‑3 human breast cancer cell line and T47D human breast cancer cell line. PVDF membrane was probed with 1 µg/mL of Rabbit Anti-Human B7‑H4 Monoclonal Antibody (Catalog # MAB65764) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for B7‑H4 at approximately 50-80 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

B7‑H4 was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Rabbit Anti-Human B7‑H4 Monoclonal Antibody (Catalog # MAB65764) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). 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 cell surfaces in cancer cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

• Reactivity: Hu
• Applications: WB, Flow, IHC, CyTOF-ready
• Clone: 2318A
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated

B7-H4 Antibody (2318A) [Unconjugated] Summary

• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: Mouse myeloma cell line, NS0-derived human B7-H4
  Phe29-Ser258
  Accession # Q7Z7D3
• Specificity: Detects human B7-H4 in direct ELISAs.
• Source: N/A
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit

Applications/Dilutions

• Dilutions:
  • Western Blot 1 ug/mL
  • Flow Cytometry 0.25 ug/10^6 cells
  • Immunohistochemistry 3-25 ug/mL
  • CyTOF-ready

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for B7-H4 Antibody (2318A) [Unconjugated]

• 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

Background

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).

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Uniprot:
  • Q7Z7D3()