Source	Human embryonic kidney cell, HEK293-derived cynomolgus monkey VISTA/B7-H5/PD-1H protein Phe33-Ala194, with a C-terminal 6-His tag
Purity	>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin level	<0.10 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis	Phe33
Predicted Molecular Mass	20 kDa
SDS-PAGE	35-45 kDa, under reducing conditions
Activity	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.

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

	Recombinant Cynomolgus VISTA/B7-H5/PD-1H His-tag Protein Binding Activity 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.
	Recombinant Cynomolgus VISTA/B7-H5/PD-1H His-tag Protein SDS-PAGE 2 μg/lane of Recombinant Cynomolgus Monkey VISTA/B7-H5/PD-1H His-tag Protein (Catalog # 10473-B7) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 35-45 kDa.

Formulation, Preparation and Storage

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

10473-B7

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution	Reconstitute at 200 μg/mL in PBS.
Shipping	The product is shipped at ambient temperature. Upon receipt, store it 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. 3 months, -20 to -70 °C under sterile conditions after reconstitution.

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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Background: VISTA/B7-H5/PD-1H

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).

Item	Value
References	Flajnik, M.F. et al. (2012) Immunogenetics 64:571. Wilcox, R.A. et al. (2012) Eur. J. Haematol. 88:465. Nowak EC et al. (2017) Immunol Rev. 276:66. Sakr, M.A. et al. (2010) Cancer Sci. 101:2368. Aloia, L. et al. (2010) J. Biol. Chem. 285:7776. Parisi, S. et al. (2012) FASEB J. 26:3957. Wang, L. et al. (2011) J. Exp. Med. 208:577. Mehta N et al. (2019) Cell Rep. 28:2509 Wang J. et al. (2019) Immunology, 156:74. Johnston R.J. et al. (2019) Nature 574:565.
Entrez Gene IDs	64115 (Human); 74048 (Mouse); 690899 (Rat); 102116468 (Cynomolgus Monkey)
Alternate Names	4632428N05Rik, B7-H5, B7H5, C10orf54, Dies1, Gi24, PD-1H, PD1H, PP2135, SISP1, VISTA, VSIR, chromosome 10 open reading frame 54, platelet receptor Gi24, stress induced secreted protein 1

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

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