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BV421 Mouse Anti-Human TIGIT
BV421 Mouse Anti-Human TIGIT

Multicolor flow cytometric analysis of TIGIT expression on peripheral blood lymphocytes. Whole blood was costained with Alexa Fluor®647 Mouse Anti-Human CD56 (Cat. No. 557711), Alexa Fluor®488 Mouse Anti-Human CD4 (Cat. No. 557695), BUV395 Mouse Anti-Human CD45RO (Cat. No. 564291 or 564292), and either BD Horizon™ BV421 Mouse IgG2b, κ Isotype Control (Cat. No. 562748, Top Plots) or BD OptiBuild™ BV421 Mouse Anti-Human TIGIT (Cat. No. 747844, Bottom Plots) at 0.5 μg/test. After staining, the cells were treated with 1× BD Pharm Lyse™ Lysing Solution (Cat. No. 555899) to remove erythrocytes.

     Left Plots: The pseudocolor dot plots showing the correlated expression of TIGIT [or Ig Isotype control staining] versus CD56 were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes.

     Right Plots: The pseudocolor dot plots showing the coexpression of TIGIT [or Ig Isotype control staining] versus CD45RO were derived from gated viable CD4-positive lymphocytes.

Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

The above is qualification data only and does not represent a specific OptiBuild™ lot.

Multicolor flow cytometric analysis of TIGIT expression on peripheral blood lymphocytes. Whole blood was costained with Alexa Fluor®647 Mouse Anti-Human CD56 (Cat. No. 557711), Alexa Fluor®488 Mouse Anti-Human CD4 (Cat. No. 557695), BUV395 Mouse Anti-Human CD45RO (Cat. No. 564291 or 564292), and either BD Horizon™ BV421 Mouse IgG2b, κ Isotype Control (Cat. No. 562748, Top Plots) or BD OptiBuild™ BV421 Mouse Anti-Human TIGIT (Cat. No. 747844, Bottom Plots) at 0.5 μg/test. After staining, the cells were treated with 1× BD Pharm Lyse™ Lysing Solution (Cat. No. 555899) to remove erythrocytes.

     Left Plots: The pseudocolor dot plots showing the correlated expression of TIGIT [or Ig Isotype control staining] versus CD56 were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes.

     Right Plots: The pseudocolor dot plots showing the coexpression of TIGIT [or Ig Isotype control staining] versus CD45RO were derived from gated viable CD4-positive lymphocytes.

Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

The above is qualification data only and does not represent a specific OptiBuild™ lot.

Product Details
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BD OptiBuild™
TIGIT; T-cell immunoreceptor with Ig and ITIM domains; VSIG9; VSTM3; WUCAM
Human (Tested in Development)
Mouse IgG2b, κ
Human TIGIT Recombinant Protein
Flow cytometry (Qualified)
0.2 mg/ml
AB_2872307
Aqueous buffered solution containing ≤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 the dye under optimum conditions that minimize unconjugated dye and antibody.

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

  1. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
  2. Researchers should determine the optimal concentration of this reagent for their individual applications.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. 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.
  7. 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.
  8. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  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.
747844 Rev. 2
Antibody Details
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741182

The 741182 monoclonal antibody specifically recognizes TIGIT (T cell Immunoreceptor with Ig and ITIM domains) which is also known as Vstm3 (V-set and transmembrane domain-containing 3), Vsig9 (V-set and Ig domain-containing 9) and WUCAM (Washington University Cell Adhesion Molecule). TIGIT is a 30-34 kDa single-pass type I transmembrane glycoprotein that belongs to the CD28 family within the Ig superfamily. TIGIT has an extracellular region with a V-type Ig-like domain, transmembrane sequence, and a cytoplasmic domain with an immunoreceptor tyrosine-based inhibitory motif (ITIM). TIGIT is expressed on NK cells and subsets of activated and memory T cells, regulatory T cells (Treg), and T follicular helper (Tfh) cells. TIGIT binds to CD112 (PVRL2/Nectin-2) and CD155 (PVR/Necl-5) that are expressed on dendritic cells (DC), endothelial cells, fibroblasts, and some tumor cells and can induce IL-10 release and inhibition of IL-12 production. Ligand-bound TIGIT downregulates TCR-mediated T cell activation and proliferation, and can block NK cell-mediated cytotoxicity.

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 BlueTM 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.

747844 Rev. 2
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.
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BV421
Violet 405 nm
407 nm
423 nm
747844 Rev.2
Citations & References
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Development References (3)

  1. Downs-Canner S, Berkey S, Delgoffe GM, et al. Suppressive IL-17A(+)Foxp3(+) and ex-Th17 IL-17A(neg)Foxp3(+) Treg cells are a source of tumour-associated Treg cells.. Nat Commun. 2017; 8:14649. (Clone-specific: Flow cytometry). View Reference
  2. Yasuma K, Yasunaga J, Takemoto K, et al. HTLV-1 bZIP Factor Impairs Anti-viral Immunity by Inducing Co-inhibitory Molecule, T Cell Immunoglobulin and ITIM Domain (TIGIT).. PLoS Pathog. 2016; 12(1):e1005372. (Clone-specific: Flow cytometry). View Reference
  3. Yu X, Harden K, Gonzalez LC, Francesco M, et al. The surface protein TIGIT suppresses T cell activation by promoting the generation of mature immunoregulatory dendritic cells. Nat Immunol. 2009; 10(1):48-57. (Biology). View Reference
747844 Rev. 2

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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.