Tcrγδ

Manufactured by Thermo Fisher Scientific

Sourced in Germany

The TCRγδ is a laboratory instrument used for the detection and analysis of T cell receptor (TCR) γδ cells. It provides a tool for researchers to identify and quantify the presence of this specific subtype of T cells in biological samples.

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Lab products found in correlation

Cd163, by Thermo Fisher Scientific (2 mentions) Live dead fixable blue viability dye, by Thermo Fisher Scientific (2 mentions) Lsr fortessa 1 flow cytometer, by BD (2 mentions) Cd16 cd32 93, by BioLegend (2 mentions) Ifn γ, by Thermo Fisher Scientific (2 mentions) Il 17a, by BioLegend (2 mentions) Facsdiva, by BD (2 mentions) F4 80, by BioLegend (2 mentions) Siglec f, by BD (2 mentions) Cd206, by BioLegend (2 mentions) Mast cell tryptase, by Agilent Technologies (1 mentions) Dc sign, by Abcam (1 mentions) Impress detection system, by Vector Laboratories (1 mentions) Il 17a, by R&D Systems (1 mentions) Permount, by Thermo Fisher Scientific (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Cd11b, by Thermo Fisher Scientific (1 mentions) Collagenase 4, by Merck Group (1 mentions) Dnase 1, by Merck Group (1 mentions) Intracellular fixation and permeabilization buffer kit, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

TCRγδ (GL3, (4 citations) TCRγδ(5A6.E9) (3 citations) Biotin-TCRγδ (2 citations) Anti-TCRγδ (2 citations) TCRγδ-PE (2 citations)

7 protocols using tcrγδ

Multiparameter Flow Cytometry Protocol

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Fluorescently labelled antibodies purchased from eBioscience (Waltham, MA) include the following: TCRβ (biotin; 1:200), TCRγδ (biotin; 1:200), and CD19 (biotin; 1:200), TCRβ (H57-597; 1:200), CD19 (eBio1D3; 1:200), TCRγδ (EbioGL3; 1:200), CD45.2 (104; 1:400), CD163 (TNKUPJ; 1:100) and IFNγ (XMG1.2; 1:100). Antibodies purchased from Biolegend (San Diego, CA) include the following: CD16/CD32 (93; 1:500), Streptavidin, CD11b(M1/70; 1:500), F4/80 (BM8; 1:200), CD206 (C068C2; 1:200), and IL-17A (TC11-18H10.1, 1:100). Antibodies purchased from BD Pharmingen (Billerica, MA) include the following: Siglec-F (E50-2440; 1:800), CD64 (X54-5/7.1; 1:200) and CD4 (RM4.5; 1:200). Antibodies purchased from Life Technologies (Washington, DC) include the following: LIVE/DEAD Fixable Blue Viability Dye (1:500). Cells were collected on an LSR Fortessa I flow cytometer equipped with FACSDIVA (BD Biosciences) software and data were analyzed with FlowJo software (Tree Star, Ashland, OR).

Karmele E.P., Pasricha T.S., Ramalingam T.R., Thompson R.W., Gieseck RL I.I.I., Knilans K.J., Hegen M., Farmer M., Jin F., Kleinman A., Hinds D.A., Pereira T.A., de Queiroz Prado R., Bing N., Tchistiakova L., Kasaian M.T., Wynn T.A, & Vannella K.M. (2019). Anti-IL-13Rα2 therapy promotes recovery in a murine model of inflammatory bowel disease. Mucosal immunology, 12(5), 1174-1186.

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Multiparameter Flow Cytometry Protocol

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Immunohistochemical Profiling of Tissue Samples

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Formalin-fixed tissues were embedded in paraffin and sectioned into 5-μm sections, deparaffinized and rehydrated, followed by heat-induced epitope retrieval. Methanol containing 3% H₂O₂ was used to block the endogenous peroxidase for 15-30 minutes. Sections were then blocked with BSA and incubated for 1h at room temperature or overnight at 4°C with primary antibody to CD3, CD138, CD68, DC-SIGN (Abcam), IL-17A (R&D Systems), mast cell tryptase (Dako), CD15 (Novus), CD56 (Thermo Scientific), TCRγδ (eBioscience) or isotype controls. After washing with PBS three times, immunolabeling was performed using the ImPress detection system (Vector) followed by visualization with ImmPACT DAB peroxidase substrate. Finally, the specimens were counterstained with Mayer’s hematoxylin, mounted with Permount (Fisher), and sections were scanned using a ScanScope (Aperio).

Moutsopoulos N., Konkel J., Sarmadi M., Eskan M.A., Wild T., Dutzan N., Abusleme L., Zenobia C., Hosur K.B., Abe T., Uzel G., Chen W., Chavakis T., Holland S.M, & Hajishengallis G. (2014). Defective Neutrophil Recruitment in Leukocyte Adhesion Deficiency Type I Disease Causes Local IL-17-driven Inflammatory Bone Loss. Science translational medicine, 6(229), 229ra40.

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Comprehensive Immune Cell Profiling

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Spleen and lymph nodes cells were isolated by mechanical desegregation in PBS+3% FBS. Whole skin was minced and digested 3hrs at 37°C in DMEM (Gibco) with 1mg/ml collagenase-IV (Sigma) and 1mg/ml DNase I (Sigma) and strained. Cell suspensions were stained using antibodies from EBioscience: TCR-β (H57), CD4 (RM4.5), CD8 (53-6.7), CD19 (Ebio1D3), CD90.2 (53-2.1), CD45 (30F11), CD44 (IM7), NK1.1 (PK136), TCR-γδ (EbioGL3), CD11c (N418), CD11b (M1/70), IL-33R (ST2), CD25 (PC61, 7D4), and Foxp3 (FJK16s). Foxp3 staining was performed using the eBioscience kit. Cells were acquired on a LSR II (BD Biosciences) and analyzed with FlowJo.

Grinberg-Bleyer Y., Dainichi T., Oh H., Heise N., Klein U., Schmid R.M., Hayden M.S, & Ghosh S. (2015). NF-κB p65 and c-Rel control epidermal development and immune homeostasis in the skin. Journal of immunology (Baltimore, Md. : 1950), 194(6), 2472-2476.

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Multiparameter Flow Cytometry Analysis

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Cell suspensions were obtained from intestines, spleens, and MLNs and stained in PBS 2% FCS with conjugated antibodies to the following markers: TCR-β (eBioscience), TCR-γδ (eBioscience), CD3 (eBioscience), CD45 (eBioscience), CD4 (eBioscience), CD8a (eBioscience), CD62L (eBioscience), CD44 (BD), CD45R (B220; eBioscience), CD357 (GITR; eBioscience), CCR9 (eBioscience), α4β7 (BioLegend), Foxp3 (eBioscience), IgA (eBioscience), IgM (eBioscience), CD25 (eBioscience), CD103 (eBioscience), CD45RB (BioLegend), and Nrp-1 (R&D Systems). Live/dead cell discrimination was obtained using the Aqua Dead Cell Stain kit (Invitrogen). For cytokine production, cells were stimulated for 4 h with 20 ng/ml PMA (Sigma-Aldrich) and 0.5 µg/ml ionomycin (Sigma-Aldrich). Golgi Stop (1,000×; BD) was added during the last 3 h of stimulation. Cells were fixed and permeabilized using intracellular fixation and permeabilization buffer kit (eBioscience) and stained for conjugated anti–IL-17A (eBioscience) and anti–IFN-γ (eBioscience). Flow cytometry data were acquired at FACSCanto II. Data were analyzed with FlowJo software (version 7.6.5).

Rigoni R., Fontana E., Guglielmetti S., Fosso B., D’Erchia A.M., Maina V., Taverniti V., Castiello M.C., Mantero S., Pacchiana G., Musio S., Pedotti R., Selmi C., Mora J.R., Pesole G., Vezzoni P., Poliani P.L., Grassi F., Villa A, & Cassani B. (2016). Intestinal microbiota sustains inflammation and autoimmunity induced by hypomorphic RAG defects. The Journal of Experimental Medicine, 213(3), 355-375.

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Lineage Cocktail for ILC2 Identification

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For the lineage cocktail, biotin-coupled antibodies against B220, CD4, CD8, CD11b, CD11c, CD19, FcϵRI, Gr-1, NK1.1, TCRβ, TCRγδ, Ter119 and SiglecF (eBioscience) were used. Fluorescent coupled streptavidin or antibodies against CD45, ST2, CD90⁺, human CD2, CD25, KLRG1 and CD127 (Biolegend) were used.
For intracellular staining antibodies against IL-5 (BD Pharmingen), IL-13 (eBioscience) and GATA-3 (eBioscience) were used. To determine the cell viability, we stained the cells using Zombie aqua reagent (Biolegend). The ILC2s were sorted using a Facs ARIA II sorter in the Instituto Nacional de Enfermedades Respiratorias (INER) and the Instituto de Investigaciones Biomédicas, UNAM. The samples were analyzed using an Attune Nxt cytometer (Thermofisher) located in the Laboratorio Nacional de Citometría de Flujo of Instituto de Investigaciones Biomédicas, UNAM and a BD FACSMelody Cell Sorter located in the Instituto de Fisiología Celular.

Olguín-Martínez E., Muñoz-Paleta O., Ruiz-Medina B.E., Ramos-Balderas J.L., Licona-Limón I, & Licona-Limón P. (2022). IL-33 and the PKA Pathway Regulate ILC2 Populations Expressing IL-9 and ST2. Frontiers in Immunology, 13, 787713.

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Intestinal TCRαβ and TCRγδ Immunostaining

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Frozen intestinal tissues were cut into 10 μm sections with a frozing microtome (Leica, CM1950, Germany). After blocked with 5% normal goat serum, frozen sections were incubated with FITC-labelled TCRαβ or TCRγδ (eBioscience) antibodies in the dark at 4 C for 3 h, and finally counterstained with DAPI for observation by fluorescence microscope (Zeiss AX10, Carl Zeiss AG, Germany).

Zhang X., Zhu B., Li L., Xu J., Han Y., Zhang J, & Hua Z. (2022). The dephosphorylation of FADD at S191 induces an excessive expansion of TCRαβ⁺ IELs in the intestinal mucosa. Immunology, 167(2).

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