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Alexa Fluor® 647 Mouse Anti-Human VISTA
Alexa Fluor® 647 Mouse Anti-Human VISTA
Multiparameter flow cytometric analysis of VISTA expression on human peripheral blood leucocyte populations. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leucocytes were then washed and stained with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 565571; Left Plot) or Alexa Fluor® 647 Mouse Anti-Human VISTA antibody (Cat. No. 566670/566671; Right Plot). Two-parameter flow cytometric contour plots showing the correlated expression of VISTA (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of viable leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Multiparameter flow cytometric analysis of VISTA expression on human peripheral blood leucocyte populations. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leucocytes were then washed and stained with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 565571; Left Plot) or Alexa Fluor® 647 Mouse Anti-Human VISTA antibody (Cat. No. 566670/566671; Right Plot). Two-parameter flow cytometric contour plots showing the correlated expression of VISTA (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of viable leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
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BD Pharmingen™
VSIR; B7-H5; B7H5; C10orf54; DD1alpha; GI24; SISP1
Human (QC Testing)
Mouse IgG1, κ
Human VISTA Transfected Cell Line
Flow cytometry (Routinely Tested)
5 µl
AB_2744493
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 to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  6. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  7. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566671 Rev. 1
Antibody Details
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MIH65.rMAb

The MIH65.rMAb is a recombinant monoclonal antibody derived from MIH65 hybridoma cells. MIH65.rMAb specifically binds to human V-domain Ig suppressor of T cell activation (VISTA) like the conventional MIH65 antibody and performs like the MIH65 antibody when used to stain cells and analyze them by flow cytometry. VISTA is also known as B7-H5 (B7H5), PD-1H, DD1alpha (DD1α), chromosome 10 open reading frame 54 (C10orf54), Platelet Receptor Gi24 (GI24), or Stress induced secreted protein 1 (SISP1). VISTA is a 55-65 kDa single-pass, type I transmembrane glycoprotein that is encoded by VSIR (V-set immunoregulatory receptor) which belongs to the V-set domain containing family within the Ig superfamily. VISTA is primarily expressed on precursors and mature cells of the hematopoietic lineage including, monocytes, macrophages, neutrophils, dendritic cells (DC), T cells, regulatory T cells (Treg), and natural killer (NK) cells but not B cells. VISTA functions as an inhibitory regulator that can suppress T cell proliferation and cytokine production.

            

566671 Rev. 1
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
566671 Rev.1
Citations & References
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Development References (2)

  1. Bharaj P, Chahar HS, Alozie OK, et al. Characterization of programmed death-1 homologue-1 (PD-1H) expression and function in normal and HIV infected individuals.. PLoS ONE. 2014; 9(10):e109103. (Biology). View Reference
  2. 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
566671 Rev. 1

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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

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