Recombinant Cynomolgus VISTA/B7-H5/PD-1H His-tag Protein, CF

When Recombinant Human VSIG3 Fc Chimera (Catalog # 9229-VS) is present at 0.5 μg/mL (100 μL/well), the concentration of Recombinant Cynomolgus Monkey VISTA/B7-H5/PD1-H His-tag (Catalog # 10473-B7) that produces a 50% optimal binding response is found to be 75-375 ng/mL.

2 μg/lane of Recombinant Cynomolgus Monkey VISTA/B7-H5/PD-1H His-tag Protein (Catalog # 10473-BL) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at kDa.

• Reactivity: Pm-Cm
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Cynomolgus VISTA/B7-H5/PD-1H His-tag Protein, CF Summary

• Details of Functionality: Measured by its binding ability in a functional ELISA. When Recombinant Human VSIG3 Fc Chimera (Catalog # 9229-VS) is represent at 0.5 µg/mL (100 µL/well), the concentration of Recombinant Cynomolgus Monkey VISTA/B7-H5/PD-1H His-tag (Catalog # 10473-B7) that produces a 50% optimal binding response is found to be 75-375 ng/mL.
• Source: Human embryonic kidney cell, HEK293-derived cynomolgus monkey VISTA/B7-H5/PD-1H protein
  Phe33-Ala194, with a C-terminal 6-His tag
• Accession #: XP_005565644.1
• N-terminal Sequence: Phe33
• Protein/Peptide Type: Recombinant Proteins
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 20 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 35-45 kDa, under reducing conditions

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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 200 μg/mL in PBS.

Notes

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

Alternate Names for Recombinant Cynomolgus VISTA/B7-H5/PD-1H His-tag Protein, CF

• 4632428N05Rik
• B7H5
• B7-H5
• C10orf54
• chromosome 10 open reading frame 54
• Dies1
• Gi24
• PD1H
• PD-1H
• platelet receptor Gi24
• PP2135
• SISP1
• stress induced secreted protein 1
• VISTA
• VSIR

Background

V-type immunoglobulin domain-containing suppressor of T-cell activation (VISTA), also known as B7-H5, Platelet receptor Gi24, and SISP1, is a transmembrane glycoprotein with homology to B7 family of immune co‑stimulatory molecules (1, 2). Mature cynomolgus VISTA consists of an extracellular domain (ECD) with one V‑type Ig‑like domain, a transmembrane segment, and a cytoplasmic domain containing a Src homology 2 (SH2)-binding motif and three C-terminal SH3-binding regions (3). Within the ECD, cynomolgus VISTA shares 96% and 69% amino acid sequence identity with human and mouse VISTA, respectively. In human, the ECD can be shed by MT1‑MMP, leaving behind a fragment in the membrane (4). VISTA promotes both MT1‑MMP expression and the MT1‑MMP mediated activation of MMP2 (4). VISTA supports the differentiation of embryonic stem cells (ESC) and enhances BMP4 induced signaling in ESC but is also down‑regulated following BMP4 exposure (5, 6). VISTA can bind to BMP4 directly and is also capable of associating with the type I BMP receptor Activin RIB/ALK4 (5, 6). VISTA is expressed on the surface of naïve CD4+ T cells and regulatory T cells and functions as an immune checkpoint protein involved in the regulation of T cell activity (3, 7, 8). VISTA is up‑regulated in vivo on activated monocytes and dendritic cells and its overexpression results in reduced CD4+ and CD8+ T cell activation and proliferation and decreased IL2 and IFN-gamma production (6, 7). VISTA expression on tumor cells attenuates the anti‑tumor immune response and enables more rapid tumor progression (7). In contrast, VISTA limits disease progression in the autoimmune disease model EAE (7). VISTA has recently shown to interact with VSIG3 and PSGL1 (9, 10).

1. Flajnik, M.F. et al. (2012) Immunogenetics 64:571.
2. Wilcox, R.A. et al. (2012) Eur. J. Haematol. 88:465.
3. Nowak EC et al. (2017) Immunol Rev. 276:66.
4. Sakr, M.A. et al. (2010) Cancer Sci. 101:2368.
5. Aloia, L. et al. (2010) J. Biol. Chem. 285:7776.
6. Parisi, S. et al. (2012) FASEB J. 26:3957.
7. Wang, L. et al. (2011) J. Exp. Med. 208:577.
8. Mehta N et al. (2019) Cell Rep. 28:2509
9. Wang J. et al. (2019) Immunology, 156:74.
10. Johnston R.J. et al. (2019) Nature 574:565.

Bioinformatics

• Uniprot:
  • XP_005565644.1()