Anti tcrβ pe cy7

Manufactured by BioLegend

Sourced in United States

Anti-TCRβ-PE-Cy7 is a fluorochrome-conjugated monoclonal antibody that recognizes the T cell receptor beta chain (TCRβ) on the surface of T cells. This antibody can be used for the identification and characterization of T cell populations in flow cytometry applications.

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Anti-TCRβ (12 citations) TCRβ-Pe Cy7 (5 citations) Anti-TCRβ-PE (3 citations) TCRβ–PE (2 citations)

6 protocols using anti tcrβ pe cy7

Multicolor Flow Cytometry for Immune Cell Profiling

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PE anti-CD3, Pacific Blue anti-CD8α, biotin-conjugated anti-CD8β, PE Cy7 anti-CD4, APC Cy7 anti-CD62L, APC anti-CD103, PerCP Cy5.5 anti-TCRγδ, PE Cy7 anti-TCRβ, APC Cy7-B220, PerCP Cy5.5 anti-B220, FITC-EpCAM, PE Cy7 anti-CD11b, APC anti-F4/80, biotin-conjugated anti-IgA antibodies were purchased from BioLegend. Streptavidin-PE was purchased from BD Biosciences.

Ito K., Nakajima A., Fukushima Y., Suzuki K., Sakamoto K., Hamazaki Y., Ogasawara K., Minato N, & Hattori M. (2017). The potential role of Osteopontin in the maintenance of commensal bacteria homeostasis in the intestine. PLoS ONE, 12(3), e0173629.

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Spectrum Flow Cytometry of tdTomato+ BAMs

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Spectrum flow cytometry was conducted to assess the frequency of tdTomato⁺ BAMs.^{79 (link),80 (link)} Mice were euthanized and intracardiac perfused with cold PBS to remove blood. Dura mater, enriched cortex, and choroid plexus were collected, and single cell suspension was prepared according to the published protocol.^{76 (link)} Single cell suspensions incubated with a combination of the following antibodies along with a Fc blocking antibody: Ghost Dye Red 780 (1:1 000, Tonbo Biosciences, 13–0865), APC/Fire810 anti-NK1.1 (1:100, BioLegend, 156519), BB700 anti-MHC II (1:500, BD Biosciences, 746197), APC anti-Ly6G (1:100, BD Biosciences, 560599), PE-CF594 anti-CD45 (1:1000, BD Biosciences, 562420), PE-Cy5 anti-CD11b (1:1000, Tonbo Biosciences, 55–0112-U100), PE-Cy7 anti-TCRβ (1:100, Biolegend, 109221), Spark NIR 685 anti-B220 (1:200, BioLegend, 103267).
Samples were then assessed by the spectral flow cytometer (Cytek Aurora, Cytek Biosciences) equipped with SpectroFlo software (Cytek Biosciences). Acquired files were then analyzed by FlowJo software (BD Life Sciences).

Wang L., Zheng J., Zhao S., Wan Y., Wang M., Bosco D.B., Kuan C.Y., Richardson J.R, & Wu L.J. (2024). CCR2+ monocytes replenish border-associated macrophages in the diseased mouse brain. Cell reports, 43(4), 114120.

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Single-cell RNA-seq of Regulatory T Cells

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Splenocytes from Foxp3^YFP-creIcos^+/+ (ICOS WT) and Foxp3^YFP-creIcos^fl/fl (ICOS FC) male mice were isolated 6 dpi with NP-OVA/alum and stained with anti-CD4 Alexa Fluor 647 (GK1.5; BioLegend), anti-TCRβ PE-Cy7 (H57-597; BioLegend), and propidium iodide (Thermo Fisher Scientific). Live (PI⁻), conventional (YFP⁻), and regulatory (YFP⁺) CD4⁺TCRβ⁺ T cells were sorted with a BD FACSAria (BD Biosciences) to >95% purity. Sorted conventional and regulatory T cells were mixed in a 1:10 ratio to provide an internal control. A total of 13,500 cells from ICOS WT and ICOS FC mice were sent for library preparation. Libraries were generated using the following components from 10×Genomics: Chromium Next GEM Chip G Single Cell Kit, Chromium Next GEM Single Cell 3′ GEM, Library & Gel Bead Kit v3.1, and Chromium i7 Multiplex Kit. Sequencing was performed by Genome Québec using a NovaSeq 6000 (Illumina) with a flow cell S1 PE28*91.

Panneton V., Mindt B.C., Bouklouch Y., Bouchard A., Mohammaei S., Chang J., Diamantopoulos N., Witalis M., Li J., Stancescu A., Bradley J.E., Randall T.D., Fritz J.H, & Suh W.K. (2023). ICOS costimulation is indispensable for the differentiation of T follicular regulatory cells. Life Science Alliance, 6(4), e202201615.

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Multiparameter Flow Cytometry Panels

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The following antibodies (BioLegend, San Diego, CA, USA) were added to 100 µL aliquots of 10⁵ Fc-blocked splenocytes prepared as described above to make a final 1/600 dilution: anti-CD1d-PE(clone 1B1), anti-B220-FITC (clone RA3-6B2), anti-CD93-APC (clone AA4.1), anti-CD138-PE-Cy7 (clone 281-2), anti-GL7-PE (clone GL7), anti-Ly6G-Alexa488 (clone 1A8), anti-Ly6C-PE (clone HK 1.4), anti-CD4-FITC (clone GK 1.5), anti-CD8-PE (clone 53-6.7), anti-NK1.1-APC(clone PK 136), anti-TCRβ-PE-Cy7 (clone H57-597).

Radwanska M., Nguyen H.T, & Magez S. (2021). African Trypanosomosis Obliterates DTPa Vaccine-Induced Functional Memory So That Post-Treatment Bordetella pertussis Challenge Fails to Trigger a Protective Recall Response. Vaccines, 9(6), 603.

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Multiparameter Flow Cytometry Panels

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Magez S., Li Z., Nguyen H.T., Pinto Torres J.E., Van Wielendaele P., Radwanska M., Began J., Zoll S, & Sterckx Y.G. (2021). The History of Anti-Trypanosome Vaccine Development Shows That Highly Immunogenic and Exposed Pathogen-Derived Antigens Are Not Necessarily Good Target Candidates: Enolase and ISG75 as Examples. Pathogens, 10(8), 1050.

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Multiparametric Flow Cytometry Analysis

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For surface staining, cells were washed in PBS and stained with Live/Dead Fixable Blue Dead Cell Stain (ThermoFisher Scientific) for 25 min on ice. Cells were washed and incubated for 25 min on ice with anti-CD25-BV421 (BioLegend, San Diego, CA, USA), anti-CD44-PE (BioLegend), anti CD4-PE-TexasRed (ThermoFisher Scientific), anti-CD8-PerCP-Cy5.5 (BioLegend), anti-TCRβ-PE-Cy7 (BioLegend), and anti-CD45RB-FITC (BioLegend). Cells were washed, fixed in 1% (v/v) methanol-free formaldehyde, and analyzed on an LSRII (BD Biosciences).
For intracellular staining, after staining with Live/Dead Fixable Blue Dead Cell Stain and surface antibodies as described above, cells were washed and fixed with 2% formaldehyde (v/v) for 15 min on ice. Fixed cells were washed and permeabilized with PBS/1% BSA supplemented with 0.03% saponin for 10 min on ice. Cells were washed and incubated with anti-cleaved caspase-3 Alexa 647 (Cell Signaling) for 30 min on ice. Cells were washed, fixed in 1% formaldehyde (v/v), and analyzed on an LSRII (BD Biosciences).
All flow cytometry data were analyzed with FlowJo v10 software (FlowJo, Ashland, OR, USA).

Secinaro M.A., Fortner K.A., Dienz O., Logan A., Murphy M.P., Anathy V., Boyson J.E, & Budd R.C. (2018). Glycolysis promotes caspase-3 activation in lipid rafts in T cells. Cell Death & Disease, 9(2), 62.

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