Flow cytometric analysis of human whole blood cells stained with VISTA (D1L2G™) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) and co-stained with CD19-APC and CD14-APC. VISTA is present on CD14+ Monocytes (bottom right; upper right quadrant) and shows an increase in intensity when compared to the isotype control (bottom left; upper left quadrant). CD19+ B cells are negative for VISTA (top right; upper left quadrant) and do not show an increase in intensity when compared to the isotype control (top left; upper left quadrant).
This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 488 fluorescent dye and tested in-house for direct flow cytometric analysis in human cells. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated VISTA (D1L2G™) XP® Rabbit mAb #64953.
Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: If using whole blood, lyse red blood cells and wash by centrifugation prior to fixation.
posted July 2009
revised June 2017
Protocol Id: 407
VISTA (D1L2G™) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) recognizes endogenous levels of total VISTA protein.Species Reactivity:
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human VISTA protein.
VISTA (V-Domain Ig Suppressor of T Cell Activation) is a negative checkpoint control protein that regulates T cell activation and immune responses. VISTA, which contains a single Ig-like V-type domain, a transmembrane domain, and an intracellular domain, has sequence similarity to both the B7 and CD28 family members. Although primarily expressed by myeloid cells, VISTA is also expressed by CD4+, CD8+, and FoxP3+ T-cells. Thus, VISTA is described as both a ligand and a receptor (1-3). Blocking VISTA induces T-cell activation and proliferation, and potentiates disease severity in the EAE model (1). Furthermore, genetic deletion of VISTA in mice leads to spontaneous T-cell activation and chronic inflammation (4,5). In mouse models of cancer, neutralization of VISTA enhances T-cell proliferation and effector function and increases tumor infiltration, suggesting VISTA blockade could be an effective strategy for tumor immunotherapy (6,7).
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Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
D1L2G is a trademark of Cell Signaling Technology. Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
The Alexa Fluor dye conjugates in this product are sold under license from Life Technologies Corporation, for research use only excluding use in combination with DNA microarrays and high content screening (HCS).
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