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.

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.

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.

Applications for Human/Mouse B7-H4 Antibody

Application

Recommended Usage

CyTOF-ready

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

Flow Cytometry

0.25 µg/10⁶ cells
Sample: HEK293 Human Cell Line Transfected with Mouse B7-H4 and eGFP

Immunohistochemistry

5-25 µg/mL
Sample: Perfusion fixed frozen sections of mouse kidney

Western Blot

1 µg/mL
Sample: Mouse uterus tissue, Mouse placenta tissue, and JEG‑3 human epithelial choriocarcinoma cell line

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

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

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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

Yi, K.H. and L. Chen (2009) Immunol. Rev. 229:145.
Salceda, S. et al. (2005) Exp. Cell Res. 306:128.
Zang, X. et al. (2003) Proc. Natl. Acad. Sci. 100:10388.
Prasad, V.R. et al. (2003) Immunity 18:863.
Sica, G.L. et al. (2003) Immunity 18:849.
Kryczek, I. et al. (2006) J. Exp. Med. 203:871.
Tringler, B. et al. (2005) Clin. Cancer Res. 11:1842.
Xue, Q. et al. (2010) Stem Cells Dev. 19:27.
Song, H. et al. (2008) Cancer Lett. 266:227.
Park, G.B. et al. (2009) Immunology 128:360.
Zang, X. et al. (2007) Proc. Natl. Acad. Sci. 104:19458.
Krambeck, A.E. et al. (2006) Proc. Natl. Acad. Sci. 103:10391.
Simon, I. et al. (2006) Cancer Res. 66:1570.
Thompson, R.H. et al. (2008) Cancer Res. 68:6054.
Azuma, T. et al. (2009) PloS Med. 6:e1000166.
Suh, W.-K. et al. (2006) Mol. Cell. Biol. 26:6403.

Long Name

B7 Homolog 4

Alternate Names

B7H4, B7S1, B7x, Vtcn1

Entrez Gene IDs

79679 (Human); 242122 (Mouse); 295322 (Rat)

Gene Symbol

VTCN1

UniProt

Q7TSP5

Additional B7-H4 Products

Product Documents for Human/Mouse B7-H4 Antibody

Product Datasheets

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COA

SDS

Download SDS

Product Specific Notices for Human/Mouse B7-H4 Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human/Mouse B7-H4 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB21541

Key Product Details

Species Reactivity

Applications

Label

Antibody Source

View all B7-H4 Antibodies »

Product Specifications

Immunogen

Specificity

Clonality

Host

Isotype

Scientific Data Images for Human/Mouse B7-H4 Antibody

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