Anti tcrβ

Manufactured by BioLegend

Anti-TCRβ is a lab equipment product that detects the T cell receptor beta chain. It is a tool used for the identification and analysis of T cells in various research applications.

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Lab products found in correlation

Anti cd4, by BioLegend (7 mentions) Anti cd45, by BioLegend (7 mentions) Anti b220, by BioLegend (5 mentions) Anti cd11b, by BioLegend (4 mentions) Anti cd44, by BioLegend (4 mentions) Anti cd19, by BioLegend (3 mentions) Anti t bet, by BioLegend (3 mentions) Anti cd8α, by BioLegend (2 mentions) Anti cd11c, by BioLegend (2 mentions) Anti cd3, by BioLegend (2 mentions) Anti cd73, by BioLegend (2 mentions) Anti cd11c, by Thermo Fisher Scientific (2 mentions) Anti cd19, by BD (2 mentions) Anti cd169, by BioLegend (2 mentions) Anti cd138, by BioLegend (2 mentions) Anti cd62l, by BioLegend (2 mentions) Live dead fixable near ir dead cell stain kit, by Thermo Fisher Scientific (2 mentions) Trizol reagent, by Thermo Fisher Scientific (2 mentions) Anti b220 cd45r, by BioLegend (1 mentions) Anti 1 a 1 e, by BioLegend (1 mentions)

Spelling variants of this product

Anti-TCRβ (H57-597, (8 citations) Anti-TCRβ-FITC (6 citations) Anti-TCRβ-PE-Cy7 (6 citations) Anti-TCRβ (clone H57-597, (6 citations) Anti-mouse TCRβ (5 citations) Anti-TCRβ-BV421 (3 citations) Anti-TCRβ antibody (H57-597) (2 citations) Anti-TCRβ chain and anti-TRAIL (CD253) (2 citations) APC anti-mouse TCRβ chain (clone H57-597), (2 citations) Anti-TCRβ antibody (2 citations)

12 protocols using anti tcrβ

Multiparametric Flow Cytometry for Immune Cell Analysis

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To determine chimerism, splenocytes were stained with anti‐H‐2D^d‐FITC (clone: 34–2–12) and anti‐H‐2D^b‐PE (clone: KH95) (all from Biolegend). Treg cells were determined in the spleens according to manufacturer's instructions using a kit from eBioscience (San Diego, CA). Spleen cells were also characterized using anti‐CD11b (clone: M1/70), anti‐CD11c (clone: N418), anti‐B220/CD45R (clone: RA3‐6B2), anti‐I‐A/I‐E (clone: M5/114.152), anti‐CD4 (clone: RM4‐5 and GK1.5), anti‐pDCA1 (clone: 927), anti‐CD8α (clone: 100712), anti‐TCR‐β (clone: H57‐597), all from Biolegend, and a viability marker (LIVE/DEAD^® Fixable Near‐IR Dead Cell Stain Kit, Life Technologies, Eugene, OR). For MACS cell purification, we used anti‐CD11c (cat. 130‐052‐001), anti‐CD4 (cat. 130‐049‐201), and anti‐CD8 (cat. 130‐049‐301) microbeads (Miltenyi Biotec Bergisch Glabach, Germany). For the in vitro functional assays, DC subpopulations were purified from spleens by MACS. CD11c cells were then sorted by FACS (BD FacsAria III) using APC/Cy7‐labeled anti‐B220, PercP‐labeled anti‐I‐A/I‐E, APC‐labeled anti‐pDCA1, PE‐labeled anti‐CD11b and PE/Cy7‐CD8a. We obtained 90.9 ± 1.35% purity for CD8αcDCs, 95.1 ± 1% for CD11b cDCs and 94.± 0.3% for pDCs. All cells were acquired using a FACS‐LSRFortessa according to BD bioscience protocols and analyzed by FlowJo software version 9.8.1.

Gaignage M., Marillier R.G., Uyttenhove C., Dauguet N., Saxena A., Ryffel B., Michiels T., Coutelier J, & Van Snick J. (2017). Mouse nidovirus LDV infection alleviates graft versus host disease and induces type I IFN‐dependent inhibition of dendritic cells and allo‐responsive T cells. Immunity, Inflammation and Disease, 5(2), 200-213.

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Comprehensive Immune Cell Profiling

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Surface staining of the single cell suspensions was performed for 30 min at 4°C, and viability was assessed using LIVE/DEAD Fixable viability dye as per the manufacturer’s instructions (Thermo Fisher Scientific).
The following surface markers antibodies from Biolegend were used: anti-CD3 (clone: 145–2C11), anti-CD4 (RM4–5), anti-CD8a (53–5.8), anti-CD45 (30-F11), anti-CD11b (M1/70), anti-CD11c (N418), anti-CD19 (6D5), anti-TCRβ (H57–597), anti-NK1.1 (PK136), anti-KLRG1 (2F1), anti-PD-1 (29F.1A12), anti-CD25 (PC-61), anti-CD44 (IM7), anti-CD62L (MEL-14), anti-Thy1.1 (OX-7), anti-CCR2 (SA203G11), anti-CXCR3 (CXCR3–173), anti-CXCR5 (L138D7), anti-CXCR6 (SA051D1), anti-CD103 (2E7). Samples were then fixed overnight at 4°C using the using 100 μL of Foxp3 Fix/Perm buffer (eBioscience). After membrane permeabilization using 1X permeabilization buffer (eBioscience) for 5 min, intracellular staining was performed for 120 min at room temperature using the following antibodies: anti-Ctla-4 (UC10–4B9, Biolegend), anti-Helios (22F6, Biolegend), anti-Foxp3 (FJK16, Thermofisher), anti-Gata3 (TWAJ, Thermofisher), anti-RORγ (AFKJS-9, Thermofisher), anti-Ki-67 (16A8, Biolegend). Cells were acquired with an Aurora flow cytometer (Cytek Biosciences) or a FACSymphony flow cytometer (BD Biosciences). Data were analyzed using FlowJo software version 10 (TreeStar, BD LifeSciences).

Leon J., Chowdhary K., Zhang W., Ramirez R.N., André I., Hur S., Mathis D, & Benoist C. (2023). Mutations from patients with IPEX ported to mice reveal different patterns of FoxP3 and Treg dysfunction. Cell reports, 42(8), 113018.

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Murine Immune Cell Profiling

Cited 1 time

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A single cell suspension from thymi and spleens of 4–6 week-old mice was obtained using a Dounce homogenizer. Cells were first preincubated for 10 min on ice with Fc receptor-blocking anti-CD16/32 (clone 93) antibody in PBS-1% biotin-free BSA solution and then stained for 20 min with specific primary antibodies. Biotinylated primary antibodies were revealed with streptavidin conjugated fluorescent dyes (SAv-PE/Cy7, SAv-APC, SAv-APC/Cy7). Labeled cells were analyzed on BD FACSCantoII (BD Biosciences) or CytoFLEX (Beckman Coulter) flow cytometers and data analysis was performed using FlowJo software (FlowJo, LLC). Clones of monoclonal antibodies were: anti-CCR6 (29–2L17), anti-CD3ε (145–2C11), anti-CD4 (GK1.5), anti-CD5 (53–7.3), anti-CD8 (53–6.7), anti-CD11c (N418), CD24 (M1/69), anti-CD25 (PC61), anti-CD27 (LG.3A10), anti-CD28 (E18), anti-CD44 (IM7), anti-CD45.1 (A20), anti-CD45.2 (104), anti-CD71 (RI7217), anti-CD73 (TY/11.8), anti-CD117 (c-Kit, 2B8) anti-B220 (RA3–6B2), anti-CD11b (Mac1, M1/70), anti-γδT-cell (GL3), hamster IgG-PE/Cy7 (HTK888), anti-Ly6G/Ly6C (GR1, RB6–8C5), anti-Nk1.1 (PK136), anti-Tcrβ (H57–597), anti-Tcrβ(Vα2) (B20.1), Ter-119 (all from Biolegend). Splenocytes were analyzed as previously described (18 (link)).

Zikmund T., Kokavec J., Turkova T., Savvulidi F., Paszekova H., Vodenkova S., Sedlacek R., Skoultchi A.I, & Stopka T. (2019). ISWI ATPase Smarca5 Regulates Differentiation of Thymocytes Undergoing β-selection.. Journal of immunology (Baltimore, Md. : 1950), 202(12), 3434-3446.

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Immunofluorescent Staining of Mouse Spleen Sections

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7 mm spleen sections were prepared from OCT-frozen tissues, fixed in acetone for 10 min, and stored at −80°C. The slides were thawed and re-hydrated using PBS and blocked using PBS+1% BSA and 2.4G2 for 10 minutes. The slides were then stained with relevant Abs as described in the figure legends in a dark humid chamber for 30 minutes. The Abs were either purified in our lab or purchased and are as follows. Anti-B220 (Biolegend RA3-6B2), anti-CD19 (BD ID3), anti-IgM (home-made B7-6), for B cells, anti-CD4 (Biolegend GK1.5), anti-TCR-β (Biolegend H57-597) for T cells, anti-CD138 (Biolegend 281-2) for PB, anti-CD169 (Biolegend 3D6.112) for metalophillic macrophages, anti-CD11c (ebioscience N418) and anti-T-bet (Biolegend 4B10). The slides were washed thrice using PBS and the same steps were followed for secondary Ab. For intracellular staining, the sections were permeabilized using 0.3% Triton X-100 reagent before staining. Sections were mounted using ProLong Anti-fade Gold (Life Technologies) and imaged using Olympus Fluorescence Microscope IX3-BSW and acquired using Cell Sens Dimension software.

Trivedi N., Weisel F., Smita S., Joachim S., Kader M., Radhakrishnan A., Clouser C., Rosenfeld A., Chikina M., Vigneault F., Hershberg U., Ismail N, & Shlomchik M. (2019). Liver is a generative site for the B cell response to Ehrlichia muris. Immunity, 51(6), 1088-1101.e5.

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Splenocyte T- and B-cell Profiling

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Flow cytometry analysis was performed to assess the presence of T- and B-cells in splenocytes from the wt and TCRα knock-out mice. Briefly, spleens were harvested, homogenised and then the homogenate was collected and pelleted. Red blood cells in the pellet were lysed and the cells were washed, then resuspended in FACS buffer (PBS +2% fetal bovine serum + 2mM EDTA) and counted. Cells (10⁶ cell/ml) were stained with anti-CD4, anti-TCRβ or anti-B220 antibodies (all from Biolegend; 1:400 dilution) and then analysed by flow cytometry using an Accuri Flow Cytometer (BD). 10,000 events were assessed per sample and the data was analysed using FlowJo software.

Stone V.M., Utorova R., Butrym M., Sioofy-Khojine A.B., Hankaniemi M.M., Ringqvist E.E., Blanter M., Parajuli A., Pincikova T., Fischler B., Karpati F., Hytönen V.P., Hyöty H., Hjelte L, & Flodström-Tullberg M. (2022). Coxsackievirus B infections are common in Cystic Fibrosis and experimental evidence supports protection by vaccination. iScience, 25(10), 105070.

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Multiparametric Flow Cytometry of B Cells

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For FC staining, non-specific binding was blocked using anti-FcR clone 2.4G2 and dead cells were excluded using cell viability dye (Tonbo Biosciences). The Abs were either purified in our lab or purchased and are as follows. Anti-B220 (Biolegend, RA3-6B2), anti-CD19 (BD ID3), anti-IgM (homemade, B7-6), anti-CD45 (home-made 30-F11), anti-CD21 (homemade, 7G6), anti-CD23 (ebioscience, B3B4) for B cells, anti-CD4 (Biolegend, GK1.5), anti-TCR-β (Biolegend H57-597) for T cells, anti-CD138 (Biolegend, 281-2) and anti-CD44 (Biolegend, IM7) for B cell blasts and PB, anti-CD73 (Biolegend, TY-11.8), anti-CD80 (ebioscience, 16-1oA1), anti-PD-L2 (ebioscience, TY25) for MBC, PNA (vector labs), anti-CD95 (BD, Jo2) for GCBC, anti-CD169 (Biolegend, 3D6.112) for metalophillic macrophages, anti-CD11b (Biolegend M1-70), anti-CD11c (ebioscience, N418), anti-CD69 (ebioscience, H1.2F3), anti-AID (ebioscience, mAID-2) and anti-T-bet (Biolegend, 4B10). The click IT Plus Edu kit was purchased from Invitrogen and the staining was done according to the recommended protocol. The cells were analyzed on LSR II or Fortessa instruments (BD) and the data were analyzed on FlowJo software.

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Flow Cytometry Antibody Panel for Immune Cell Profiling

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The following anti‐mouse monoclonal antibodies for flow cytometry were purchased from Thermo Fisher Scientific, BD Biosciences, or Biolegend: anti‐TCRβ (H57‐597, cat# 109224, 109206), anti‐CD62L (MEL‐14, cat# 746726, 104441, 17‐0621‐83), anti‐CD69 (H1.2F3, cat# 564684, 612793), anti‐CD122 (TM‐β1, cat# 123207, 123216), anti‐CD4 (GK1.5, cat# 11‐0041‐85), anti‐CD8a (53‐6.7, cat# 100712 anti‐CD44 (IM7, cat# 11‐0441‐85, 103030, 56‐0441‐82), anti‐NK1.1 (PK136, cat#564144), anti‐Ly49C/F/I/H (14B11, cat# 108209), (90 cat#56‐0381‐82), anti‐CD39 (24DM51, cat# 25‐0391‐82), anti‐CD49a (Ha31/8, cat# 562113), anti‐CXCR6 (SA051D1, cat# 151109), anti‐P2 × 7 (1F11 cat# 94464), anti‐CCR7 (4B12, cat# 562675) anti‐TCF7 (S33‐966, cat# 564217), anti‐KI67 (B56, cat# 561277), anti‐CD3Ɛ (145‐2C11; cat# 563565), anti‐T‐bet (eBio4B10, cat# 12‐5825‐82), anti‐RORγt (Q31‐378, cat# 564722), GATA3 (L50‐823, cat# 560068), anti‐CD45.1 (A20, cat# 110714), and anti‐CD45.2 (104, cat# 109822). Dead cells were excluded from our analysis using the LIVE/ DEAD Fixable Near‐IR Dead Cell Stain kit (Invitrogen; L10119, cat# 2134021). To detect iNKT cells, tetramers of CD1d containing the α‐GalCer derivative PBS57 (cat# 44870) were obtained from the Tetramer Core Facility of the US National Institutes of Health.

Kragten N.A., Taggenbrock R.L., Parga Vidal L., van Lier R.A., Stark R, & van Gisbergen K.P. (2022). Hobit and Blimp‐1 instruct the differentiation of iNKT cells into resident‐phenotype lymphocytes after lineage commitment. European Journal of Immunology, 52(3), 389-403.

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T Cell Apoptosis Evaluation by Flow

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T lymphocytes were incubated with an FcR-blocking antibody (2.4G2), stained with FITC-, PE-, PE/Cy5-, APC-, APC-Cy7-, or Pacific Blue-labeled mAbs on ice for 20 min, and washed with FACS buffer (2% FBS, 0.02% NaN₃ in PBS). Then cells were re-suspended in Annexin V-binding buffer (10 mM HEPES, pH 7.4, 140 mM NaCl, 2.5 mM CaCl₂) and incubated with Annexin V-PE (BD Bioscience) and 7-AAD (BD Bioscience) at room temperature for 15 min. The cells were then diluted in Annexin V-binding buffer and analyzed by flow cytometry within one hour. A total of 0.5–20×10⁵ events were collected on a FACSCanto II flow cytometer (BD Biosciences) and analyzed using FlowJo software (Tree Star). All fluorescence-labeled Abs, including anti-CD3, anti-CD4, anti-CD8, anti-TCRβ, anti-CD19 were obtained from BioLegend.

He M.X, & He Y.W. (2015). c- FLIP protects T lymphocyte from apoptosis in the intrinsic pathway. Journal of immunology (Baltimore, Md. : 1950), 194(7), 3444-3451.

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Comprehensive Immunophenotyping of T Cells

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Cell culture media was RPMI 1640 (Mediatech Inc.) supplemented with 10% heat-inactivated FBS, 100 U/ml penicillin, 100 mg/ml streptomycin, 2 mM L-glutamine, and 50 mM 2-mercaptoethanol (Sigma-Aldrich). Fluorescent anti-CD4, anti-CD8α, anti-CD8β, anti-CD25, anti-CD44, anti-CD45, anti-CD45.1, anti-CD45.2, anti-CD62L, anti-CD103, anti-CCR4, anti-CCR6, anti-CCR7, anti-CCR-9, anti-TCRβ and anti-TCRγδ antibodies were purchased from Biolegend. Anti-CTLA-4, anti-PD-1, anti-GITR, anti-CD127, anti-IFNγ, anti-CXCR3 and anti-CXCR4 were purchased from BD Biosciences-Pharmingen. Anti-MHCII, anti-CD207, anti-Gr-1, anti-CD11b, and anti-Foxp3 staining kit were purchased from eBioscience. Murine recombinant IL-2, IL-6 and E-selection-FC were purchased from R&D Systems. BD Cytofix/Perm buffer was used for intracellular staining. Cells were run on a BD LSR II flow cytometer or a Beckman Coulter Gallios flow cytometer and analyzed by Flowjo (Flowjo LLC).

Zhang R., Borges C.M., Fan M.Y., Harris J.E, & Turka L.A. (2015). Requirement for CD28 in Effector Treg Differentiation, CCR6 induction, and Skin Homing. Journal of immunology (Baltimore, Md. : 1950), 195(9), 4154-4161.

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Hematopoietic Stem Cell Reconstitution

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SJL (CD45.1) mice were irradiated at 475cGly rad twice (950cGly in total) at least 2 hours apart. Donor BM cells (CD45.2), together with competitor BM cells (SJL) (3 × 10⁵/each), were injected into the tail veins of SJL recipients. Peripheral blood samples from recipients were periodically analyzed to assess frequencies of donor blood cells, including myeloid, B, and T cells. Red blood cells were lysed with RBC lysis buffer (Biolegend) before antibody staining. The antibodies used to analyze donor chimerism were anti-CD45.1, anti-CD45.2, anti-CD11b, anti-B220, and anti-TCR-β (all from Biolegend).

Hirata Y., Furuhashi K., Ishii H., Li H.W., Pinho S., Ding L., Robson S.C., Frenette P.S, & Fujisaki J. (2018). CD150high bone marrow Tregs maintain hematopoietic stem cell quiescence and immune privilege via adenosine. Cell stem cell, 22(3), 445-453.e5.

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