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BD Horizon™ BV421 Mouse Anti-Human VISTA
Clone MIH65.rMAb (also known as MIH65) (RUO)
Multiparameter flow cytometric analysis of VISTA expression on human peripheral blood leucocyte populations. Human whole blood was stained with either BD Horizon BV421™ Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or BD Horizon BV421 Mouse Anti-Human VISTA antibody (Cat. No. 566750/566751; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899).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 cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
Multiparameter flow cytometric analysis of VISTA expression on human peripheral blood leucocyte populations. Human whole blood was stained with either BD Horizon BV421™ Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or BD Horizon BV421 Mouse Anti-Human VISTA antibody (Cat. No. 566750/566751; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899).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 cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
Multiparameter flow cytometric analysis of VISTA expression on human peripheral blood leucocyte populations. Human whole blood was stained with either BD Horizon BV421™ Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or BD Horizon BV421 Mouse Anti-Human VISTA antibody (Cat. No. 566750/566751; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899).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 cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
Multiparameter flow cytometric analysis of VISTA expression on human peripheral blood leucocyte populations. Human whole blood was stained with either BD Horizon BV421™ Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or BD Horizon BV421 Mouse Anti-Human VISTA antibody (Cat. No. 566750/566751; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899).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 cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
Product Details
BD Horizon™
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_2869848
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
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome-conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. 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
- 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).
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
Companion Products
Stain Buffer (FBS) RUO
Size
500 ML
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 ML
Cat No.
554657
Brilliant Stain Buffer RUO
Size
1000 Tests
Cat No.
566349
Brilliant Stain Buffer RUO
Size
100 Tests
Cat No.
563794
Brilliant Stain Buffer Plus RUO
Size
1000 Tests
Cat No.
566385
Lysing Buffer RUO
Size
100 ML
Cat No.
555899
566751 Rev. 2
Antibody Details
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.
The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max near 407 nm and Em Max near 421 nm, BD Horizon BV421 can be excited by the violet laser (405 nm) and detected with a 450/50 nm filter. BD Horizon BV421 conjugates are very bright, often exhibiting a 10-fold improvement in brightness compared to Pacific Blue™ conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon BV421, Pacific Blue, and BD Horizon V450 cannot be used simultaneously.
566751 Rev. 2
Format Details
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.
BV421
407 nm
423 nm
566751 Rev.2
Citations & References
View product citations for antibody "566751" on CiteAb
Development References (2)
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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
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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
566751 Rev. 2
Please refer to Support Documents for Quality Certificates
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.