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PE Mouse Anti-Human TIGIT
PE Mouse Anti-Human TIGIT
Multicolor flow cytometric analysis of TIGIT expression on peripheral blood leucocytes. Whole blood was stained with Alexa Fluor™ 647 Mouse Anti-Human CD56 (Cat. No. 563443; Middle Plots), BD Horizon™ BV421 Mouse Anti-Human CD4 (Cat. No. 562424), and BD Horizon™ BUV395 Mouse Anti-Human CD45RO (Cat. No. 564291/564292; Right Plots) antibodies, and with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Top Plots) or PE Mouse Anti-Human TIGIT antibody (Cat. No. 568672/568673; Bottom Plots) as indicated. Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.      Left Plots: The pseudocolor density plot showing the correlated expression of TIGIT (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of viable leucocyte populations.      Middle Plots: The pseudocolor density plot showing the correlated expression of TIGIT (or Ig Isotype control staining) versus CD56 was derived from gated events with the light scatter characteristics of viable lymphocytes.      Right Plots: The pseudocolor density plot showing the correlated expression of TIGIT (or Ig Isotype control staining) versus CD45RO was derived from CD4-positive gated events with the light scatter characteristics of viable lymphocytes.
Multicolor flow cytometric analysis of TIGIT expression on peripheral blood leucocytes. Whole blood was stained with Alexa Fluor™ 647 Mouse Anti-Human CD56 (Cat. No. 563443; Middle Plots), BD Horizon™ BV421 Mouse Anti-Human CD4 (Cat. No. 562424), and BD Horizon™ BUV395 Mouse Anti-Human CD45RO (Cat. No. 564291/564292; Right Plots) antibodies, and with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Top Plots) or PE Mouse Anti-Human TIGIT antibody (Cat. No. 568672/568673; Bottom Plots) as indicated. Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.      Left Plots: The pseudocolor density plot showing the correlated expression of TIGIT (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of viable leucocyte populations.      Middle Plots: The pseudocolor density plot showing the correlated expression of TIGIT (or Ig Isotype control staining) versus CD56 was derived from gated events with the light scatter characteristics of viable lymphocytes.      Right Plots: The pseudocolor density plot showing the correlated expression of TIGIT (or Ig Isotype control staining) versus CD45RO was derived from CD4-positive gated events with the light scatter characteristics of viable lymphocytes.
Product Details
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BD Pharmingen™
T-cell immunoreceptor with Ig and ITIM domains; VSIG9; VSTM3; WUCAM
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human TIGIT Transfected Cell Line
Flow cytometry (Routinely Tested)
5 µl
201633
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 the dye under optimum conditions and unconjugated antibody and free dye were removed.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation).  When fluorochrome conjugated antibodies are bound to BD® 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 cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  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. 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).
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. An isotype control should be used at the same concentration as the antibody of interest.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
568673 Rev. 1
Antibody Details
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TgMab-2

The TgMab-2 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.

568673 Rev. 1
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
568673 Rev.1
Citations & References
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Development References (3)

  1. Chiang EY, Mellman I. TIGIT-CD226-PVR axis: advancing immune checkpoint blockade for cancer immunotherapy.. J Immunother Cancer. 2022; 10(4):e004711. (Biology). View Reference
  2. Shibuya A, Shibuya K. DNAM-1 versus TIGIT: competitive roles in tumor immunity and inflammatory responses.. Int Immunol. 2021; 33(12):687-692. (Biology). 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
568673 Rev. 1

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