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Two-color flow cytometric analysis of VISTA expression on mouse splenocytes. Mouse splenic leucocytes were stained with PE Rat Anti-Mouse CD8a antibody (Cat. No. 553033/553032/561095) and either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602: Left Plot) or BD Horizon BV421 Rat Anti-Mouse VISTA antibody (Cat. No. 566271/566272; Right Plot). Two-color flow cytometric contour plots showing the correlated expression of CD8a versus VISTA (or Ig isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable splenocytes. Flow cytometric analysis was performed using a BD™ Canto II Flow Cytometry System.
BD Horizon™ BV421 Rat Anti-Mouse VISTA
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The MIH64 monoclonal antibody specifically binds to V-domain Ig suppressor of T cell activation (VISTA) which is also known as, Differentiation of ESCs 1 (Dies1). VISTA is a type I transmembrane glycoprotein that belongs to the Ig superfamily. VISTA has homology to the B7 family ligand, PD-L1 (CD274), and is likewise known as PD-1H. VISTA is expressed on naïve and activated T cells, natural killer (NK) cells, macrophages, dendritic cells, and neutrophils. VISTA functions as an inhibitory ligand. It can serve as a negative immune-checkpoint protein that suppresses T cell cytokine production and proliferation. It is also involved in regulating stem cell differentiation.
Development References (5)
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Flies DB, Han X, Higuchi T, et al. Coinhibitory receptor PD-1H preferentially suppresses CD4⁺ T cell-mediated immunity.. J Clin Invest. 2014; 124(5):1966-75. (Biology). View Reference
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Flies DB, Wang S, Xu H, Chen L. Cutting edge: A monoclonal antibody specific for the programmed death-1 homolog prevents graft-versus-host disease in mouse models.. J Immunol. 2011; 187(4):1537-41. (Biology). View Reference
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Kondo Y, Ohno T, Azuma M. Effects of anti-VISTA mAb and its combination therapy in antitumor immunity. Abstract.. J Immunol. 2015; 194(1 Supplement):69.32. (Clone-specific: In vivo exacerbation). View Reference
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Ohno T, Kondo Y, Azuma M. T cell-dependent and-independent regulatory roles of a new inhibitory molecule VISTA. J Immunol. 2015; 194(1 Supplement):113.16. (Clone-specific: In vivo exacerbation).
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Wang L, Rubinstein R, Lines JL, et al. VISTA, a novel mouse Ig superfamily ligand that negatively regulates T cell responses.. J Exp Med. 2011; 208(3):577-92. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.