RB780 Mouse Anti-Human TIGIT

BD Horizon™ RB780 Mouse Anti-Human TIGIT

Name: RB780 Mouse Anti-Human TIGIT
Brand: RB780 Mouse Anti-Human TIGIT
SKU: 569940

Clone TgMab-2 (RUO)

Multicolor flow cytometric analysis of TIGIT expression on Human peripheral blood leukocytes. Fresh whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. After washing, the leukocytes were preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219/564220) and then stained with APC Mouse Anti-Human CD56 (NCAM-1) [Cat. No. 555518; Middle Plots], BD Horizon™ BUV395 Mouse Anti-Human CD4 (Cat. No. 564724), and BD Horizon™ BV421 Mouse Anti-Human CD45RO (Cat. No. 562641/562649; Right Plots) antibodies, and with either BD Horizon™ RB780 Mouse IgG1, κ Isotype Control (Cat. No. 568532; Top Plots) or BD Horizon™ RB780 Mouse Anti-Human TIGIT antibody (Cat. No. 569864/569940; Bottom Plots) as indicated. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 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 leukocyte populations.

Middle Plots: The pseudocolor density plot showing the correlated expression of TIGIT (or Ig Isotype control staining) versus CD56 (NCAM-1) was derived from gated events with the forward and side 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 forward and side light-scatter characteristics of viable lymphocytes.

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Product Details

Brand: BD Horizon™

Alternative Name: T-cell immunoreceptor with Ig and ITIM domains; VSIG9; VSTM3; WUCAM

Reactivity: Human (QC Testing)

Isotype: Mouse BALB/c IgG1, κ

Immunogen: Human TIGIT Transfected Cell Line

Application: Flow cytometry (Routinely Tested)

Vol. Per Test: 5 µl/test

Entrez Gene ID: 201633

RRID: AB_3685375

Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.

Regulatory Status: RUO

Preparation And Storage

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 unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

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 cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
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.
Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
For U.S. patents that may apply, see bd.com/patents.

Companion Products

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

Human BD Fc Block™ RUO

Size 50 µg Cat No. 564219

RB780 Mouse IgG1, κ Isotype Control RUO

Size 50 µg Cat No. 568532

APC Mouse Anti-Human CD56 (NCAM-1) RUO

Size 100 Tests Cat No. 555518

BV421 Mouse Anti-Human CD45RO RUO

Size 100 Tests Cat No. 562641

View Details

569940 Rev. 1

Antibody Details

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.

569940 Rev. 1

Format Details

RB780

The BD Horizon RealBlue™ 780 (RB780) Dye is part of the BD family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 781-nm. Driven by BD innovation, RB780 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB780 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-Cy7. RB780 is on average brighter than PE-Cy7 and has minimal spillover into Yellow-Green detectors.

Format: RB780

Excitation Source: Blue 488 nm

Excitation Max: 498 nm

Emission Max: 781 nm

569940 Rev.1

Citations & References

View product citations for antibody "569940" on CiteAb

Development References (3)

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. (Biology). View Reference
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. (Biology). View Reference
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

569940 Rev. 1

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.