Human/Mouse B7-H4 Antibody

Catalog # Availability Size / Price Qty
MAB21541-100
MAB21541-SP
Detection  of human and mouse B7‑H4 by Western Blot.
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Human/Mouse B7-H4 Antibody Summary

Species Reactivity
Human, Mouse
Specificity
Detects mouse B7-H4 in direct ELISAs. Detects human and mouse B7-H4 in Western blots.
Source
Recombinant Monoclonal Rabbit IgG Clone # 2319B
Purification
Protein A or G purified from cell culture supernatant
Immunogen
Mouse myeloma cell line, NS0-derived mouse B7-H4
Phe29-Pro258
Accession # Q7TSP5
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
Western Blot
1 µg/mL
See below
Flow Cytometry
0.25 µg/106 cells
See below
Immunohistochemistry
5-25 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

Western Blot Detection  of human and mouse B7‑H4 by Western Blot. View Larger

Detection of human and mouse B7‑H4 by Western Blot. 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.

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

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

Immunohistochemistry B7‑H4  in Mouse Kidney. View Larger

B7‑H4 in Mouse Kidney. 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.

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 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).

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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Cell and Tissue Staining Kits

Immunohistochemistry Reagents

Isotype Controls

Normal Rabbit IgG Control

Ctrl,CyTOF-ready

Reconstitution Buffers

Secondary Antibodies

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