B7-H4 Antibody (2319B) [Unconjugated]

HEK293 human embryonic kidney cell line transfected with either (A) mouse B7-H4 or (B) irrelevant protein and eGFP was stained with Rabbit Anti-Human/Mouse B7-H4 Monoclonal Antibody (Catalog # MAB21541) followed by APC-conjugated Goat anti-Rabbit IgG Secondary Antibody (Catalog # F0111). Quadrant markers were set based on Rabbit IgG control antibody staining (Catalog # MAB1050). View our protocol for Staining Membrane-associated Proteins.

Western blot shows lysates of mouse uterus tissue, mouse placenta tissue, and JEG‑3 human epithelial choriocarcinoma cell line. PVDF membrane was probed with 1 µg/mL of Rabbit Anti-Human/Mouse B7‑H4 Monoclonal Antibody (Catalog # MAB21541) 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 perfusion fixed frozen sections of mouse kidney using Rabbit Anti-Human/Mouse B7‑H4 Monoclonal Antibody (Catalog # MAB21541) at 10 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces of epithelial cells in convoluted tubules. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

• Reactivity: Hu, Mu
• Applications: WB, Flow, IHC, CyTOF-ready
• Clone: 2319B
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated

B7-H4 Antibody (2319B) [Unconjugated] Summary

• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: Mouse myeloma cell line, NS0-derived mouse B7-H4
  Phe29-Pro258
  Accession # Q7TSP5
• Specificity: Detects mouse B7-H4 in direct ELISAs. Detects human and mouse B7-H4 in Western blots.
• 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 5-25 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 (2319B) [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 B7x and B7S1, is a 50‑80 kDa glycosylated member of the B7 family of immune co‑stimulatory proteins (1, 2). Mature mouse B7-H4 consists of a 230 amino acid (aa) extracellular domain (ECD) with one Ig-like V-set domain and one Ig‑like C2‑set domain which is followed by a hydrophobic C‑terminal region (3‑5). Within the ECD, mouse B7‑H4 shares 90% and 99% aa sequence identity with human and rat B7‑H4, respectively. It shares 21%‑29% aa sequence identity with mouse B7‑1, B7‑2, B7‑H1, B7‑H2, B7‑H3, and PD‑L2. 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). Its binding to activated T cells dampens T cell responses and induces cell cycle arrest in the T cell (3‑5). Reverse signaling can induce either cell cycle arrest or apoptosis in the B7‑H4 expressing cell (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:
  • Q7TSP5()