Multicolor flow cytometric analysis of TIGIT expression on mouse splenocytes. Mouse splenic leucocytes were stimulated in culture (24 h) with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e (Cat. No. 553057; 10 μg/ml for coating) and soluble Purified NA/LE Hamster Anti-Mouse CD28 (Cat. No. 553294; 2 μg/ml) antibodies. The cells were harvested, washed and stained with FITC Rat Anti-Mouse CD4 (Cat. No. 553047/553046/561835; Left Panels) and PerCP-Cy™5.5 Rat Anti-Mouse CD8a (Cat. No. 551162/561109; Right Panels) antibodies, and either PE Mouse IgG1 κ Isotype Control (Cat. No. 554680; Top Panels) or PE Mouse Anti-Mouse TIGIT antibody (Cat. No. 565168; Bottom Panels). Two-color flow cytometric contour plots showing the correlated expression of CD4 or CD8a versus TIGIT (or Ig Isotype control staining) were derived from gated events with the forward and side-light scattering characteristics of viable leucocytes. Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System.
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Multicolor flow cytometric analysis of TIGIT expression on mouse splenocytes. Mouse splenic leucocytes were stimulated in culture (24 h) with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e (Cat. No. 553057; 10 μg/ml for coating) and soluble Purified NA/LE Hamster Anti-Mouse CD28 (Cat. No. 553294; 2 μg/ml) antibodies. The cells were harvested, washed and stained with FITC Rat Anti-Mouse CD4 (Cat. No. 553047/553046/561835; Left Panels) and PerCP-Cy™5.5 Rat Anti-Mouse CD8a (Cat. No. 551162/561109; Right Panels) antibodies, and either PE Mouse IgG1 κ Isotype Control (Cat. No. 554680; Top Panels) or PE Mouse Anti-Mouse TIGIT antibody (Cat. No. 565168; Bottom Panels). Two-color flow cytometric contour plots showing the correlated expression of CD4 or CD8a versus TIGIT (or Ig Isotype control staining) were derived from gated events with the forward and side-light scattering characteristics of viable leucocytes. Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System.
Product Details
BD Pharmingen™
Tigit; Vstm3; T cell immunoreceptor with Ig and ITIM domains
Mouse (QC Testing)
Mouse IgG1, κ
Mouse TIGIT Recombinant Protein
Flow cytometry (Routinely Tested)
0.2 mg/ml
100043314
AB_2739089
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 with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- Cy is a trademark of GE Healthcare.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
PE Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
554680
FITC Rat Anti-Mouse CD4 RUO
Size
0.5 mg
Cat No.
553047
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FITC Rat Anti-Mouse CD4 RUO
Size
0.1 mg
Cat No.
553046
565168 Rev. 2
Antibody Details
1G9
The 1G9 monoclonal antibody specifically binds to T cell immunoreceptor with Ig and ITIM domains (TIGIT) which is also known as, V-set and transmembrane domain-containing protein 3 (Vstm3). TIGIT is a 26 kDa, type I transmembrane glycoprotein that belongs to the poliovirus receptor (PVR) family. TIGIT is expressed on activated and memory CD4+ T cells and CD8+ T cells, follicular T helper cells, regulatory T cells, and NK cells. Its ligands include the poliovirus receptor (PVR, aka, CD155), poliovirus receptor-related protein 2 (PVRL2, aka, CD112), and poliovirus receptor-related protein 3 (PVRL3, CD113). TIGIT functions as a negative regulator of T cell and NK cell responses including proliferation, cytotoxicity, and the production of proinflammatory cytokines. TIGIT-deficient mice are reportedly more susceptible to developing autoimmune diseases in certain model systems.
565168 Rev. 2
Format Details
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.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
565168 Rev.2
Citations & References
View product citations for antibody "565168" on CiteAb
Development References (6)
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Foks AC, Ran IA, Frodermann V, Bot I et al. Agonistic anti-TIGIT treatment inhibits T cell responses in LDLr deficient mice without affecting atherosclerotic lesion development. PLoS ONE. 2013; 8(12):e83134. (Clone-specific: Flow cytometry). View Reference
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Joller N, Hafler JP, Brynedal B, et al. Cutting edge: TIGIT has T cell-intrinsic inhibitory functions. J Immunol. 2011; 186(3):1338-1342. (Immunogen: ELISA, Flow cytometry). View Reference
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Joller N, Lozano E, Burkett PR, et al.. Treg cells expressing the coinhibitory molecule TIGIT selectively inhibit proinflammatory Th1 and Th17 cell responses. Immunity. 2014; 40(4):569-581. (Biology). View Reference
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Levin SD, Taft DW, Brandt CS, et al. Vstm3 is a member of the CD28 family and an important modulator of T-cell function. Eur J Immunol. 2011; 41(4):902-915. (Biology). View Reference
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Lozano E, Dominguez Villar M, Kuchroo V, Hafler DA. Lozano E, Dominguez-Villar M, Kuchroo V, Hafler DA. J Immunol. 2012; 188(8):3869-3875. (Biology). View Reference
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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
565168 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.