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Anti t bet bv711

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Anti-T-bet-BV711 is a flow cytometry reagent that binds to the transcription factor T-bet. T-bet is a master regulator of T helper 1 (Th1) cell differentiation and is expressed in Th1 cells, natural killer (NK) cells, and group 1 innate lymphoid cells (ILC1s).

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3 protocols using anti t bet bv711

1

Comprehensive Phenotyping of Antigen-Specific T Cells

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We thawed PBMCs from cryovials and washed them as described above, and we seeded 2.5 × 106 cells/ml in RPMI 1640 with 10% FBS or 2% Phx. We stimulated PBMCs with 10 multiplicities of infection (MOIs) of strain 1100–2 fixed E. coli for 20 h and blocked extracellular transport with brefeldin A for a total of 4 h. We performed extracellular staining with fixable viability dye eFluor 780 (eBioscience), anti-CD3-BUV395 (BD Biosciences), anti-CD8-BV605 (BioLegend), anti-CD4-BV510 (BioLegend), anti-Vα7.2-PE-Cy7 (BioLegend), anti-LAG-3-BV786 (BioLegend), anti-CD25-BV650 (BioLegend), anti-PD-1-PerCPCy-5.5 (BioLegend), anti-CD161-PE/Dazzle-594 (BioLegend), anti-CD69-PE-Cy5 (Invitrogen), anti-CD161-allophycocyanin (BioLegend), and anti-human PE-MR1–5-OP-RU tetramer (NIH Tetramer Core Facility). The PBMCs were fixed and permeabilized using an Foxp3/transcription factor kit (eBioscience) and stained intracellularly with anti–granzyme B Alexa Fluor 700 (BioLegend), anti-TNF-α-eFluor-450 (Invitrogen), anti-IFN-α-FITC (BioLegend), and anti-T-bet-BV711 (BioLegend). We collected data using a five-laser LSRFortessa flow cytometer (BD Biosciences), and the flow data were analyzed using FlowJo software version 10 (BD Biosciences).
Labuz D., Cacioppo J., Li K., Aubé J, & Leung D.T. (2023). Enhancing Mucosal-Associated Invariant T Cell Function and Expansion with Human Selective Serum. ImmunoHorizons, 7(1), 116-124.
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2

Comprehensive Phenotyping of Antigen-Specific T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
We thawed PBMCs from cryovials and washed them as described above, and we seeded 2.5 × 106 cells/ml in RPMI 1640 with 10% FBS or 2% Phx. We stimulated PBMCs with 10 multiplicities of infection (MOIs) of strain 1100–2 fixed E. coli for 20 h and blocked extracellular transport with brefeldin A for a total of 4 h. We performed extracellular staining with fixable viability dye eFluor 780 (eBioscience), anti-CD3-BUV395 (BD Biosciences), anti-CD8-BV605 (BioLegend), anti-CD4-BV510 (BioLegend), anti-Vα7.2-PE-Cy7 (BioLegend), anti-LAG-3-BV786 (BioLegend), anti-CD25-BV650 (BioLegend), anti-PD-1-PerCPCy-5.5 (BioLegend), anti-CD161-PE/Dazzle-594 (BioLegend), anti-CD69-PE-Cy5 (Invitrogen), anti-CD161-allophycocyanin (BioLegend), and anti-human PE-MR1–5-OP-RU tetramer (NIH Tetramer Core Facility). The PBMCs were fixed and permeabilized using an Foxp3/transcription factor kit (eBioscience) and stained intracellularly with anti–granzyme B Alexa Fluor 700 (BioLegend), anti-TNF-α-eFluor-450 (Invitrogen), anti-IFN-α-FITC (BioLegend), and anti-T-bet-BV711 (BioLegend). We collected data using a five-laser LSRFortessa flow cytometer (BD Biosciences), and the flow data were analyzed using FlowJo software version 10 (BD Biosciences).
Bonnet G., Tyagi S., Libchaber A, & Kramer F.R. (1999). Thermodynamic basis of the enhanced specificity of structured DNA probes. Proceedings of the National Academy of Sciences of the United States of America, 96(11), 6171-6176.
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3

Comprehensive PBMC Immunophenotyping Protocol

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Peripheral blood mononuclear cells (PBMCs) were stained in four panels containing anti-CD3-PacificBlue (PacBlue) (antibodies from BD Biosciences unless otherwise indicated), anti-CD3-Alexa700, anti-CD25-PE-Cy7, anti-CD38-PE, anti-HLA-DR-PE-Cy7, anti-CCR5-APC, anti-CD123-PerCP-Cy5.5, anti-CD16-PacBlue, anti-CD80-FITC, anti-CD83-PE, anti-CD86-APC, anti-PD1-FITC, anti-PD-L1-PE, anti-HLA class I-APC, anti-CD69-APC-Cy7; anti-CD4-Qdot655, anti-CD8-PE-Cy5.5, anti-CD14-Qdot605 (Invitrogen); anti-CD45RA-ECD, anti-CD127-PE, anti-HLA-DR-ECD, anti-CD20-ECD (Beckman Coulter); anti-CD11c-Alexa700 (eBioscience); and anti-CD27-APC-Cy7 (BioLegend). A staining reagent for dead cells (Invitrogen Aqua Live/Dead Fixable Stain) was included. Cells were then washed and fixed in PBS containing 1% paraformaldehyde or permeabilized using a FOXP3 Fix/Perm kit (BioLegend) according to the manufacturer's instructions, intracellularly stained with anti-Ki67-Alexa 488 (BD Biosciences), anti-FOXP3-PacBlue, anti-T-Bet-BV711 (BioLegend), and anti-Eomes-eFluor 660 (eBioscience), fixed, and analyzed with a LSR II cytometer (Becton Dickinson) and FlowJo.
Méndez-Lagares G., Lu D., Chen C., Terrault N., Segal M.R., Khalili M., Monto A., Shen H., Manos M.M., Lanier L.L., Ryan J.C., McCune J.M, & Hartigan-O'Connor D. (2017). Memory T-cell proliferation before HCV therapy predicts antiviral immune responses and treatment success. Journal of immunology (Baltimore, Md. : 1950), 200(3), 1124-1132.
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