Femoral bone marrow, mesenteric and liver (celiac, portal) lymph nodes cells were isolated, washed, and counted. Cells were treated with FcR-block (Miltenyi Biotec), surface stains were performed and intracellular stains were carried out following the fixation-permeabilization buffer manufacturer’s protocol (eBioscience). TREGcells: anti-CD4-FITC (Miltenyi Biotec, clone GK1.5), anti-CD25-PE (eBioscience, clone PC61.5), anti-FOXP3-APC (eBioscience, clone FJK-16s). TH17 cells: anti-CD4-FITC (Miltenyi Biotec, clone GK1.5), anti-CD196-PE (Miltenyi Biotec, clone REA277), anti- RORγt-APC (Miltenyi Biotec, clone REA278). TH1 cells: anti-CD4-FITC (Miltenyi Biotec, clone GK1.5), anti-CD183-PE (Miltenyi Biotec, clone CXCR3-173), anti-T-bet-APC (Miltenyi Biotec, clone REA102). Dead cells were excluded via e450 viability dye (eBioscience). Data was acquired by the MACSQuant System. Analyses were performed via FlowJo VX software.
Anti t bet apc
Manufactured by Miltenyi Biotec
The Anti-T-bet-APC is a flow cytometry reagent that specifically binds to the T-bet transcription factor. T-bet is a master regulator of Th1 cell differentiation. This reagent can be used to identify and analyze Th1 cells in flow cytometry applications.
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Lab products found in correlation
Anti rorγt apc, by Miltenyi Biotec (2 mentions)
Anti cd183 pe, by Miltenyi Biotec (2 mentions)
Anti cd4 fitc, by Miltenyi Biotec (2 mentions)
Fcr block, by Miltenyi Biotec (2 mentions)
Anti cd25 pe, by Thermo Fisher Scientific (1 mentions)
Anti foxp3 apc, by Thermo Fisher Scientific (1 mentions)
Anti cd3 apc vio770, by Miltenyi Biotec (1 mentions)
Fixation permeabilization buffer, by Thermo Fisher Scientific (1 mentions)
Macsquant system, by Miltenyi Biotec (1 mentions)
Anti cd3 pe vio770, by Miltenyi Biotec (1 mentions)
Anti foxp3 pe, by Miltenyi Biotec (1 mentions)
2 protocols using anti t bet apc
1
Multiparametric Flow Cytometry for Immune Cell Profiling
2
Multiparametric Flow Cytometry of T Cell Subsets
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Cells were treated with FcR‐block (Miltenyi Biotec) and stained for cell surface markers. Cells were then treated with fixation/permeabilization buffer (eBioscience) to label intracellular transcription factors. TREGcells: anti‐CD3‐APC‐Vio770 (Miltenyi Biotec; clone REA641), anti‐CD4‐FITC (Miltenyi Biotec; clone REA604), anti‐CD25‐PE‐Vio770 (Miltenyi Biotec; clone 7D4), and anti‐FoxP3‐PE (Miltenyi Biotec; clone REA788). TH1 cells: anti‐CD3‐PE‐Vio770 (Miltenyi Biotec; clone REA641), anti‐CD4‐FITC (Miltenyi Biotec; clone REA604), anti‐CD183‐PE (Miltenyi Biotec; clone CXCR3‐173), anti‐T‐bet‐APC (Miltenyi Biotec; clone REA102). TH17 cells: anti‐CD3‐APC‐Vio770 (Miltenyi Biotec; clone REA641), anti‐CD4‐FITC (Miltenyi Biotec; clone REA604), anti‐RORγT‐APC (Miltenyi Biotec; clone REA278), anti‐AHR‐PE‐Vio770 (eBioscience; clone 4MEJJ). Dead cells were excluded from analysis via e450 viability dye (Invitrogen). A minimum of 10,000 gated cells were analyzed per specimen. Data were acquired by the MACSQuant System (Miltenyi Biotec) and analyzed by FlowJo 11.0 software (TreeStar, Ashland, OR, USA).
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