Tcrβ percp cy5

Manufactured by Cytek Biosciences

The TCRβ PerCp-Cy5.5 is a fluorescent-conjugated antibody used to detect and measure the expression of the T cell receptor beta chain (TCRβ) on the surface of T cells. It can be used in flow cytometry applications to identify and analyze T cell populations.

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Lab products found in correlation

Cd4 percp cy5, by Cytek Biosciences (2 mentions) Cd62l apc, by Cytek Biosciences (2 mentions) Cd44 fitc, by Cytek Biosciences (1 mentions) Cd69 pe cy7, by BioLegend (1 mentions) Cd5 percp cy5, by BioLegend (1 mentions) Cd8α pacific blue, by BioLegend (1 mentions) Tcrβ pacific blue, by BioLegend (1 mentions) Control igg, by BioXCell (1 mentions) Cd25 percp cy5, by BioLegend (1 mentions) Cd44 fitc, by BioLegend (1 mentions) Pd 1 percp cy5, by BioLegend (1 mentions) Cd4 buv395, by BD (1 mentions) Cd8 buv737, by BD (1 mentions) Cd69 fitc, by BD (1 mentions) Cd45.2 pe, by BD (1 mentions) Cd21 35, by BD (1 mentions) Cd86 percp cy5, by BD (1 mentions) Cd93 apc, by Thermo Fisher Scientific (1 mentions) Fixable viability dye efluor 780, by Thermo Fisher Scientific (1 mentions) Lsr 2, by BD (1 mentions)

3 protocols using tcrβ percp cy5

Multi-Marker Immune Cell Analysis

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The following antibodies were used for staining: CD4 PerCp-Cy5.5, TCRβ PerCp-Cy5.5, and CD62L APC (TONBO); CD44 FITC, PD-1 (29F.1A12) PE-Cy7, CD69 PE-Cy7, CD5 PerCp-Cy5.5, CD8α Pacific Blue, and TCRβ Pacific Blue (BioLegend); CD44 PE-Cy7, CD8α (APC-eFluor 780), CD4 Qdot606, and Zap70 FITC (Life Technology). In vivo anti-PD (J43) antibody and control IgG were purchased from BioXcell. For intracellular flow cytometry, antibodies against phosphorylated ERK^T202/Y204 (197G2), AKT^Thr308 (C31E5E), and ribosomal protein S6^Ser235/236 (D57.2.2E) were purchased from Cell Signaling Technology.

Hsu L.Y., Cheng D.A., Chen Y., Liang H.E, & Weiss A. (2017). Destabilizing the autoinhibitory conformation of Zap70 induces up-regulation of inhibitory receptors and T cell unresponsiveness. The Journal of Experimental Medicine, 214(3), 833-849.

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Multiparametric Flow Cytometry Analysis

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Single-cell suspensions from spleens, lymph nodes, and thymi were incubated with anti-CD16/32 at 5 µg/ml for 20 min on ice to block Fc receptors. Cells were stained with the following antibodies: CD4 PerCP-Cy55, TCRβ PerCp-Cy55, and CD62L APC (TONBO); CD44 FITC, PD-1 PerCP-Cy55, CD69 Cy7PE, CD25 PerCP-Cy55 (Biolegend); CD4 BUV395, and CD8 BUV737 (BD); Vα2 PE (phycoerythrin), Vα2 Pacific Blue, Vα2 FITC, FR4 Cy7PE, and CD73 BV605 (eBioscience); and CD25 BV605 (Life Technology). Dead cells were excluded using the live/dead fixable Near-IR death cell stain kit (Invitrogen). Intracellular Foxp3-FITC staining was done according to the manufacturer’s instructions (Life Technology). For detection of negatively selected thymocytes, caspase 3 PE (BD) was used as previously described (Breed et al., 2019 (link)). For intracellular flow cytometry, antibodies against phosphorylated ERK^T202/Y204 (mAb 197G2), AKT^S473 (Cell Signaling) were used as previously described (Hsu et al., 2009 (link)). Antibody against Alexa 647–anti-rabbit IgG was used as a secondary antibody to detect phosphorylation of ERK and AKT.

Nguyen T.T., Wang Z.E., Shen L., Schroeder A., Eckalbar W, & Weiss A. (2021). Cbl-b deficiency prevents functional but not phenotypic T cell anergy. The Journal of Experimental Medicine, 218(7), e20202477.

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Multiparameter Flow Cytometry Staining

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Cells were stained for 20 min on ice in MACS buffer (2% FCS in PBS with 1 mM EDTA) at 0.5 to 1 × 10⁶ cells per well in 96-well round-bottom plates unless otherwise specified. All mAbs were purchased from BioLegend unless otherwise indicated. The following monoclonal antibodies were used: TCRβ–BV421, TCRβ–PerCP-Cy5.5 (Tonbo), CD23–PE-Cy7, CD23–Alexa Fluor 488, CD86–Alexa Fluor 647, CD86–PerCP-Cy5.5, CD21/35–Pacific Blue, B220–BV785, CD69–FITC (BD), CD93–APC (eBioscience), CD4–BV605, CD45.2–PE, CD45.1–BV605, lambda-1 light chain–biotin (BD) (followed by streptavidin–BV605), CD19–PE, and CCR7–biotin (followed by streptavidin–BV421). CCR7-biotin was stained for 1 h at room temperature followed by streptavidin and other surface markers on ice. Dead cells were excluded using Fixable Viability Dye eFluor780 (eBioscience no. 65-0865-18). For time-course experiments all samples were plated simultaneously, collected and kept on ice, and stained in parallel. Viability for 24- to 48-h cultures was determined by pregating on B220⁺ TCRβ^--. All samples were run on a BD LSRii at 5,000 to 10,000 events per second. Flow cytometry data were analyzed using FlowJo (v10.8.0).

Wolf E.W., Howard Z.P., Duan L., Tam H., Xu Y, & Cyster J.G. (2022). GPR174 signals via Gαs to control a CD86-containing gene expression program in B cells. Proceedings of the National Academy of Sciences of the United States of America, 119(23), e2201794119.

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