Tcrαβ

Manufactured by Thermo Fisher Scientific

Sourced in United States, Germany

The TCRαβ is a laboratory equipment designed to detect and analyze T-cell receptor (TCR) alpha and beta chain expression. It provides a comprehensive solution for the identification and characterization of T-cell populations in various research applications.

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Lab products found in correlation

Permeabilization buffer, by R&D Systems (1 mentions) Anti cd1d, by BioLegend (1 mentions) Tc11 18h10, by BioLegend (1 mentions) Anti cd16 32 antibody, by BioLegend (1 mentions) Facsan, by BD (1 mentions) Il 17a, by BioLegend (1 mentions) F4 80, by Thermo Fisher Scientific (1 mentions) Zoledronic acid, by Merck Group (1 mentions) Phospho stat2 tyr690, by Cell Signaling Technology (1 mentions) Hla g, by Cell Signaling Technology (1 mentions) Hla g, by BioLegend (1 mentions) Alexa fluor 488, by BioLegend (1 mentions) β actin, by Santa Cruz Biotechnology (1 mentions) Facscanto 2 flow cytometer, by BD (1 mentions) Facsarea 2, by BD (1 mentions) Streptavidin percp cy7, by Thermo Fisher Scientific (1 mentions) Streptavidin percp cy5, by Thermo Fisher Scientific (1 mentions) Fluorescein isothiocyanate (fitc), by Thermo Fisher Scientific (1 mentions) Efluor 506, by Thermo Fisher Scientific (1 mentions) Cd45r b220, by Thermo Fisher Scientific (1 mentions)

6 protocols using tcrαβ

Flow Cytometric Immune Cell Analysis

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Antibodies employed in flow cytometric analysis were obtained from various commercial sources: anti-CD1d (Biolegend, 1B1), anti-αGalcer:CD1d (eBioscience, L363), CD86 (BD Bioscience, GL1), TCRαβ (eBioscience,IP26), F4/80 (eBioscience, BM8), IL17-A (Biolegend,TC11-18H10.1), NK1.1 (Biolegend, PK136), and CD3 (Biolegend, 17A2). Cells were blocked with anti-CD16/32 antibody (Biolegend) for 15 min before incubation with fluorescently labeled antibodies at a concentration of 2 μg/ml for 45 min on ice. Stained cells were washed once with FACS buffer and analyzed by FACSan (Becton Dickinson). For internal staining, cells stained with surface makers were fixed in 2% PFA Fixation buffer (eBioscience) at 4°C overnight, followed by 3 washes with permeabilization buffer (R&D) and incubation with permeabilization buffer for 20 min on ice before staining with internal antibody. Stained cells were washed once with permeabilization buffer and suspended in PBS for analysis. Flow cytometry data were processed with FlowJo software (Tree Star, inc.,).

Ban Y., Dong W., Zhang L., Zhou T., Altiti A.S., Ali K., Mootoo D.R., Blaho V.A., Hla T., Ren Y, & Ma X. (2019). Abrogation of Endogenous Glycolipid Antigen Presentation on Myelin-Laden Macrophages by D-Sphingosine Ameliorates the Pathogenesis of Experimental Autoimmune Encephalomyelitis. Frontiers in Immunology, 10, 404.

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Immunophenotyping of Immune Cells

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Zoledronic acid, LPS (lipopolysaccharide), puromycin, and G418 were obtained from Sigma‐Aldrich. Antibodies against human HLA‐G (sc‐21799) and β‐actin (sc‐47778) were purchased from Santa Cruz Biotechnology. PD‐L1 antibody (17952‐1‐AP) and PD‐L1 (22C3) were purchased from Proteintech and Agilent Technologies, respectively. Antibodies specific for CD3ε (NB600‐1441), PD‐L1 (FAB1561R, FAB1561G), CD4 (FAB3791R), and CD8 (NBP2‐34590AF488) were purchased from Novus Biologicals. HLA‐G (PE: #335906, Alexa Fluor 488: #335918), PD‐1 (#367412), CD3 (#317318), CD8 (#344704), CD56 (#304611), CD66b (#305104), and Vδ2 (#331418) were purchased from BioLegend. Anti‐Tyk2 (phospho‐Tyr1054/1055) (orb505746) antibody was obtained from Biorbyt. Antibodies specific for phospho‐ZAP70/Syk (Tyr319, Tyr352) (#MA5‐36963) and HLA‐G (MEM‐G/2: #MA1‐19394, 87G: MA1‐10356) were purchased from Thermo Fisher Scientific. Antibodies for CD14 (#12‐0149‐42), TCRγδ (#12‐9959‐42), TCRαβ (#11‐9955‐42), and NKG2D (#12‐5878‐42) were purchased from eBioscience. Vγ9 (#555732), hMito (ab92824), and VHH (iFluor 647: A02019, HRP: A2016) were purchased from BD Pharmingen, Abcam, and GenScript, respectively. Phospho‐Stat2 (Tyr690) (#77366) and HLA‐G (#79769) were obtained from Cell Signaling Technology. Recombinant human interleukin 2 (IL‐2) was obtained from Thermo Fisher Scientific.

Huang S., Pan C., Lin Y., Chen M., Chen Y., Jan C., Wu C., Lin F., Wang S., Lin C., Lin P., Huang W., Chiang Y., Tsai W., Chiu Y., Lin T., Chiu S, & Cho D. (2023). BiTE‐Secreting CAR‐γδT as a Dual Targeting Strategy for the Treatment of Solid Tumors. Advanced Science, 10(17), 2206856.

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Multiparametric Flow Cytometry Analysis

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Single cell suspensions from the indicated organs were stained with a combination of FITC-, PE-,-APC,-APC-Cy7 and biotin-conjugated antibodies, followed by streptavidin-PerCP-Cy5.5 or streptavidin-PerCP-Cy7 (eBioscience, San Diego, CA). The conjugated and unconjugated antibodies specific to the following antigens were purchased from BD Biosciences (San Jose, CA) and eBioscience: TCRαβ, NK1.1 (PK136), CD3ε (145-2C11), CD11c (N418), CD135/flt3 (A2F10), B220 (RA3-6B2), c-Kit (2B8), CD4 (L3T4), CD8α (53–6.72), CD11b (M1/70), CD19 (1D3), TER119, Gr-1 (RB6-8C59), FcγI/III (2.4G2), Sca-1 (E13-161.7), IL-7Rα (A7R34), CD41 (MWReg30), CD48 (HM48-1), CD150 (TC15-12F12.2), CD43 (S7), and CD34 (RAM34). Dead cells were excluded using propidium iodide (PI) or 7-AAD. Gating schemes for hematopoietic stem/progenitor cells are shown in S4 Fig. Data were collected on FACSCantoII flow cytometers (BD Bioscience) and analyzed using FlowJo software (TreeStar, Ashland, OR). A FACSAreaII (BD Bioscience) was used for cell sorting.

Yoshioka K., Oda A., Notsu C., Ohtsuka T., Kawai Y., Suzuki S., Nakamura T., Mabuchi Y., Matsuzaki Y, & Goitsuka R. (2015). Loss of the Homeodomain Transcription Factor Prep1 Perturbs Adult Hematopoiesis in the Bone Marrow. PLoS ONE, 10(8), e0136107.

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Multicolor Cytometry Analysis of Immune Cells

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Cytofluorimetry analysis was performed as previously described (16 (link)). In brief, single-cell suspensions from muscle or spleen were stained with MAbs against the following antigens: CD45 as a pan leukocyte (clone OX-1), TCRαβ (clone R7/3), CD45RA on B cells (clone OX33), CD45R/B220 on B cells (clone His24), anti-granulocytes (RP-1 and His48), CD4 (clone w3/25), CD45RC (clone OX22 or clone OX32), CD25 (clone OX39), CD8 (clone OX8), CD172a/SIRPα (clone OX41), CD161 on NK and myeloid cells (clone 3.2.3), CD163 on macrophages (clone ED2), CD68 for macrophages (clone ED1), and with viability dye eFluor506 or eFluor450 from eBiosciences to assess cell viability. Analysis was performed on a BD FACS Verse with FACSuite Software version 1.0.6. Post-acquisition analysis was performed using FlowJo software. The CD45RC⁻ population was defined using an isotype control. The CD45RC^high population was defined using as reference the levels of CD45RC expression on B cells since they always express the highest levels of CD45RC. The CD45RC^low population corresponded to the intermediate population between the CD45RC⁻ and the CD45RC^high cells.

Ouisse L.H., Remy S., Lafoux A., Larcher T., Tesson L., Chenouard V., Guillonneau C., Brusselle L., Vimond N., Rouger K., Péréon Y., Chenouard A., Gras-Le Guen C., Braudeau C., Josien R., Huchet C, & Anegon I. (2019). Immunophenotype of a Rat Model of Duchenne's Disease and Demonstration of Improved Muscle Strength After Anti-CD45RC Antibody Treatment. Frontiers in Immunology, 10, 2131.

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T-cell Activation and Cytokine Profiling

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ConA (type IV) was purchased from Sigma (St. Louis, MO, USA). Mouse T-cell enrichment columns and neutralizing antibodies against B7.1, B7.2, OX40L and CD40L were obtained from R&D Systems (Minneapolis, MN, USA). Purified antibodies against mouse CD3, CD28 and cytometric Bead Array for mouse IFN-γ were purchased from BD Biosciences. Fluorochrome-conjugated antibodies against mouse CD3, CD4, CD8, CD19, CD44, CD45.1, CD45.2, B220, NK1.1, TCRαβ and CXCR3 were obtained from eBioscience (San Diego, CA, USA). ALT Detection Kits were purchased from NanJing JianCheng Biochemical Institute (Nan Jing, Jiang Su, China).

Zhao X., Sun G., Sun X., Tian D., Liu K., Liu T., Cong M., Xu H., Li X., Shi W., Tian Y., Yao J., Guo H, & Zhang D. (2016). A novel differentiation pathway from CD4+ T cells to CD4− T cells for maintaining immune system homeostasis. Cell Death & Disease, 7(4), e2193-.

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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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