Tigit mbsa43

Manufactured by Thermo Fisher Scientific

Sourced in Germany

TIGIT (MBSA43) is a lab equipment product from Thermo Fisher Scientific. It is a targeted antibody that binds to the TIGIT (T-cell immunoreceptor with Ig and ITIM domains) protein. The core function of this product is to enable researchers to study the role of TIGIT in immune regulation and its potential as a therapeutic target.

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5 protocols using tigit mbsa43

Characterization of PBMCs by Flow Cytometry

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PBMCs were stained with the following surface and intracellular antibodies: LIVE/DEAD® Fixable Aqua Dead Cell Stain Kit, (ThermoFisher Scientific), to exclude dead cells, CD3 (SK7), CD4 (RPA-T4), CD8 (SK1 and RPA-T8), CTLA-4 (BNI3), TIM-3 (F38-2E2), CD244 (C1.7), CD71 (M-A712), LAG-3 (3DS223H), CD160 (BY55), PD-1 (EH12.1), PDL-1 (M1H1), PDL-2 (MIH18), CD11b (ICRF44), CD14 (M5E2) and Gal-9 (9M1-3) (BD Biosciences), TIGIT (MBSA43, eBioscience). Following staining cells were fixed with 4% paraformaldehyde, before analysis. Cells were acquired using an LSR FortessaSORP flow cytometer (BD Biosciences) and analysed with FlowJo software (Ashland, Oregon, USA). For CFSE labelling, PBMCs were labelled with 1.25μM CFSE (ThermoFisher Scientific) as previously described [82 (link)] before stimulation with SEB or a-CD3/CD28.

Okoye I., Namdar A., Xu L., Crux N, & Elahi S. (2017). Atorvastatin downregulates co-inhibitory receptor expression by targeting Ras-activated mTOR signalling. Oncotarget, 8(58), 98215-98232.

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Detailed Flow Cytometry Antibody Panel

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The following antibodies were used for flow cytometry. For mouse cells, CD4 (RM4-5; eBioscience), CD8a (53-6.7; BD Biosciences), CD90.2 (53-2.1, eBioscience), PD-1 (29F.1A12; BioLegend), CTLA-4 (UC10-4B9, eBioscience), CD25 (PC61; BD Biosciences), CD69 (H1.2F3; BD Biosciences), Foxp3 (FJK-16s; eBioscience), and c-Maf (sym0F1, eBioscience) were used. For human cells, CD4 (OKT4; BioLegend), CXCR4 (12G5; eBioscience), CCR7 (G043H7; BioLegend), CD127 (A019D5; BioLegend), IFN-γ (4S.B3; eBioscience), Foxp3 (236A/E7; eBioscience) CD8 (PRA-T8, BD Biosciences), CTLA4 (BNI3; BioLegend), TIGIT (MBSA43; eBioscience), CD45 (HI30, BD Biosciences), and PD-1 (MIH4; eBioscience) were used.

Seki A, & Rutz S. (2018). Optimized RNP transfection for highly efficient CRISPR/Cas9-mediated gene knockout in primary T cells. The Journal of Experimental Medicine, 215(3), 985-997.

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Multiparameter Flow Cytometry Analysis of T Cell Subsets

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We used a combination of two antibodies directed to p24: p24 KC57-PE was obtained from Beckman Coulter (6604667) and p24 28B7-APC was obtained from MediMabs (MM-0289-APC). The following antibodies were used for staining: CD3 (UCHT-1), CD4 (SK3), CD8 (RPA-T8), CD45RA (HI100), CCR7 (3D12), PD-1 (EH12.1), β1 (MAR4), β7 (FIB504), CD69 (FN50), CD25 (M-A251), HLA-DR (G46-6), CD38 (HIT2), Ki67 (MOPC-21), CD95 (DX2), CD39 (Tu66), CD127 (HIL-7R-M21), CXCR3 (1C6), CCR4 (1G1), CCR6 (11A9) were purchased from BD Bioscience. LAG-3 (FAB2319) was obtained from R&D systems, and TIGIT (MBSA43) from eBioscience. CD27 (O323), Tim-3 (F38-2E2), α4 (9F10), CD28 (CD28.2) and CD32 (FUN-2) were purchased from BioLegend. Live/Dead Aqua Cell Stain (405nm) was obtained from ThermoFisher Scientific (L34957). Detailed panels of antibodies are reported in S2 Table.

Pardons M., Baxter A.E., Massanella M., Pagliuzza A., Fromentin R., Dufour C., Leyre L., Routy J.P., Kaufmann D.E, & Chomont N. (2019). Single-cell characterization and quantification of translation-competent viral reservoirs in treated and untreated HIV infection. PLoS Pathogens, 15(2), e1007619.

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Characterization of Engineered NK-92 Cells

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NK-92, NK-92/hu14.18.28.z, and NK-92/hu14.18.28.z_RD-IL15 cells were incubated at 37 °C with K562 and EL4 target cells at an initial E/T ratio of 1:2, with fresh target cells again added to the effector cells after 24, 48, and 72 h. One day later, surface expression of marker proteins was analyzed by flow cytometry with fluorochrome-conjugated antibodies specific for NKp30 (p30-15; BD Biosciences), NKp44 (p44-8; BD Biosciences), NKp46 (9E2; Miltenyi Biotec, Bergisch Gladbach, Germany), NKG2D (BAT221; Miltenyi Biotec), NKG2A (REA110; Miltenyi Biotec), KIR2D (NKVFS1; Miltenyi Biotec), CD96 (NK92.39; Invitrogen), PD-1 (EH12.2H7; BioLegend), PD-L1 (MIH1; BD Biosciences), LAG-3 (7H2C65; BioLegend), TIGIT (MBSA43; Invitrogen), and TIM-3 (F38-2E2; BioLegend) using a FACSCanto II flow cytometer. Parental and CAR-engineered NK-92 cells cultured without target cells served as controls. Data were analyzed using FACSDiva software.

Bodden M., Häcker A., Röder J., Kiefer A., Zhang C., Bhatti A., Pfeifer Serrahima J., Ullrich E., Kühnel I, & Wels W.S. (2023). Co-Expression of an IL-15 Superagonist Facilitates Self-Enrichment of GD2-Targeted CAR-NK Cells and Mediates Potent Cell Killing in the Absence of IL-2. Cancers, 15(17), 4310.

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Multiparameter Flow Cytometry of Macaque Immune Cells

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The following fluorochrome conjugated monoclonal antibodies reactive with macaque cells were used for flow cytometry studies: CD3 (SP34-2, BD Biosciences [BD]), CD4 (L200, BD), CD8 (SK1, BD), CD95 (DX2, BD), CD28 (CD28.2, BioLegend), CCR5 (3A9, BD), CXCR5 (MU5UBEE, Invitrogen), PD-1 (EH12.2H7, BioLegend), ICOS (C398.4A, BioLegend), CCR7 (150503, BD), α4β7 (A4B7, NHP Reagent Resource), LAG3 (3DS223H, eBioscience), Tim-3 (F38-2E2, BioLegend), TIGIT (MBSA43, Invitrogen), CD20 (2H7, BioLegend), CD19 (J3-119, Beckman Coulter), HLA-DR (L243, BioLegend), CD10 (HI10A, BioLegend), CD21 (B-ly4, BD), CD27 (O323, BioLegend), IgD (IADB6, Southern Biotech), IgG (G18-145, BD), IL-21R (2G1-K12, BioLegend), Ki-67 (B56, BD), BCL6 (IG191E/A8, BioLegend), CD80 (2D10, BioLegend), CD56 (B159, BD), CD45 (D058-1283, BD), CD14 (MoP9, BD), CD16 (3G8, BD), CCR2 (48607, R&D Systems), CD11b (ICRF44, BioLegend), CD11c (3.9, Invitrogen), PD-L1 (29E.2A3, BioLegend), PD-L2 (24F.10C12, BioLegend), CD80 (2D10, BioLegend), CD86 (FUN-1, BD), CD141 (1A4, BD), CD163 (GHI/61, BioLegend), and CD123 (7G3, BD).

Kvistad D., Pallikkuth S., Sirupangi T., Pahwa R., Kizhner A., Petrovas C., Villinger F, & Pahwa S. (2021). IL-21 enhances influenza vaccine responses in aged macaques with suppressed SIV infection. JCI Insight, 6(20), e150888.

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