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BV421 Rat Anti-Mouse VISTA
BV421 Rat Anti-Mouse VISTA
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.
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.
Product Details
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BD Horizon™
Vista; PD-1H; Dies1
Mouse (QC Testing)
Rat IgG2a, κ
Mouse VISTA Transfected Cell Line
Flow cytometry (Routinely Tested)
0.2 mg/ml
387836
AB_2869745
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

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

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  6. 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.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Antibody Details
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MIH64

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.

Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
BV421
Violet 405 nm
407 nm
423 nm
Citations & References
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Development References (5)

  1. 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
  2. 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
  3. 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
  4. 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).
  5. 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
View All (5) View Less

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.