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Cd3 145 2c11

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The CD3 (145-2C11) is a lab equipment product that is used for the detection and analysis of the CD3 antigen, which is a key component of the T cell receptor complex. This product is designed to provide researchers with a reliable and accurate tool for immunological studies and flow cytometry applications.

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Lab products found in correlation

Cd4 rm4 5, by Thermo Fisher Scientific (3 mentions) Fixable viability stain 510, by BD (2 mentions) Cd62l mel 14, by BioLegend (2 mentions) Fortessa, by BD (2 mentions) Pd 1 29f 1a12, by BioLegend (2 mentions) Cd11b m1 70, by Thermo Fisher Scientific (2 mentions) Ter119, by Thermo Fisher Scientific (2 mentions) Cd25 pc61, by Thermo Fisher Scientific (2 mentions) Cd4 rm4 5, by BD (2 mentions) Facs lsr 2, by BD (2 mentions) Canto 2, by BD (2 mentions) Cd69 h1.2f3, by BD (1 mentions) P syk y348 moch1ct, by Thermo Fisher Scientific (1 mentions) Cd45.2 clone 104, by Thermo Fisher Scientific (1 mentions) Cd4 clone gk1, by Thermo Fisher Scientific (1 mentions) Cd5 clone 53 7, by Thermo Fisher Scientific (1 mentions) Igd clone 11 26, by Thermo Fisher Scientific (1 mentions) Gl 7 clone gl 7, by Thermo Fisher Scientific (1 mentions) Cd19 clone ebio1d3, by Thermo Fisher Scientific (1 mentions) Cd8 clone 53 6, by Thermo Fisher Scientific (1 mentions)

16 protocols using cd3 145 2c11

1

Multiparametric Flow Cytometry of Immune Cells

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For flow cytometry staining, dissociated cells were incubated with a fixable viability stain 510 (BD Biosciences) followed by staining with primary antibodies directed against CD45 (30-F11), CD11b (M1/70), Gr1 (RB6–8C5), F4/80 (BM8), CD206 (C068C2), MHCII (AF6–120.1), CD44 (IM7), PDL1 (MIH5), TIGIT (1G9), and CD69 (H1.2F3) obtained from BD Biosciences; CD3 (145–2C11), CD4 (GK1.5), CD8 (53–6.7), and pSykY348 (moch1ct) from eBioscience, PD1 (29F.1A12), and CD62 L (MEL-14) from BioLegend. Samples were acquired on BD Fortessa or LSRII and data were analyzed using FlowJo software version 10 (TreeStar).
Rohila D., Park I.H., Pham T.V., Weitz J., Hurtado de Mendoza T., Madheswaran S., Ishfaq M., Beaman C., Tapia E., Sun S., Patel J., Tamayo P., Lowy A.M, & Joshi S. (2023). Syk Inhibition Reprograms Tumor-Associated Macrophages and Overcomes Gemcitabine-Induced Immunosuppression in Pancreatic Ductal Adenocarcinoma. Cancer Research, 83(16), 2675-2689.
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2

Multiparameter Flow Cytometry Immunophenotyping

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CD45R/B220 (clone RA3-6B2, 1:100, eBioscience), CD3 (145-2c11, 1:100, eBioscience), CD45.2 (clone 104, 1:100, eBioscience), CD4 (clone GK1.5, 1:100, eBioscience), CD8 (clone 53-6.7, 1:100, eBioscience), CD19 (clone eBio1D3, 1:100, eBioscience), CD5 (clone 53-7.3, 1:100, eBioscience), CD21 (clone eBio4E3, 1:100, eBioscience), CD43 (clone eBioR2/60, 1:100, eBioscience), IgD (clone 11–26, 1:100, eBioscience), Gr1 (clone RB6-8C5, 1:100, eBioscience), IgM (clone 11f41, 1:100, eBioscience), GL-7 (clone GL-7, 1:100, eBioscience), Mac1 (clone M1/70, 1:100, eBioscience).
Souroullas G.P., Fedoriw Y., Staudt L.M, & Sharpless N.E. (2017). Lkb1 deletion in murine B lymphocytes promotes cell death and cancer. Experimental hematology, 51, 63-70.e1.
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3

Isolation and Analysis of Mucosal Immune Cells

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Cells from the colon and small intestine lamina propria were isolated as previously described8 (link). The cells were stimulated and stained as previously described8 (link). The following antibodies were used for surface staining of: CD3 (145-2C11, eBioscience); CD4 (L3T4, BD Difco); CD11b (M1/70, eBioscience); CD11c (N418, eBioscience); F4/80 (BM8, eBioscience); CD103 (M290, BD Difco); major histocompatibility complex (MHC) II (M5/114.15.2, BD Dico); TCR-γδ (eBioGL3, eBioscience); and NKp46 (29A1.4, eBioscience). Intracellular cytokine staining was performed using IL-17A (TC11-18H10, BD Difco) and IL-22 (IL-22JOP, eBioscience) antibodies. All antibodies were used at final concentration of 1 μg/ml. The cells were analyzed using a Gallios flow cytometer (Beckman Coulter, Brea, CA, USA). Leukocytes were gated using forward scatter (FSC) and side scatter (SSC), and within the leukocyte gates, the innate immune cells were identified as macrophages (MHCII+F4/80+CD103−CD11b+CD11c−) or dendritic cells (MHCII+F4/80− CD103+/−CD11b−CD11c+). For the lymphoid compartment, the leukocytes were gated using FSC and SSC. Within the lymphocyte gate, the populations were identified as TH17 cells (CD3+CD4+IL-17+IL-22+), TH22 cells (CD3+CD4+ IL-17−IL-22+), NKp46+ ILCs (including ILC3 and NK cells; CD3−CD4−NKp46+), LTi cells (CD3−CD4+NKp46−), γδ T cells (CD3+CD4−TCRγδ+) or CD3−CD4−NKp46− cells.
Lamas B., Richard M.L., Leducq V., Pham H.P., Michel M.L., Da Costa G., Bridonneau C., Jegou S., Hoffmann T.W., Natividad J.M., Brot L., Taleb S., Couturier-Maillard A., Nion-Larmurier I., Merabtene F., Seksik P., Bourrier A., Cosnes J., Ryffel B., Beaugerie L., Launay J.M., Langella P., Xavier R.J, & Sokol H. (2016). CARD9 impacts colitis by altering gut microbiota metabolism of tryptophan into aryl hydrocarbon receptor ligands. Nature medicine, 22(6), 598-605.
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4

Immune cell profiling in tumor samples

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Single-cell suspensions from tumors or splenocytes were incubated with anti-Fc receptor antibody (2.4G2) on ice for 15 minutes in FACS buffer (PBS with 1% FBS and sodium azide). The cells were then stained with the appropriate antibodies in 2.4G2-containing FACS buffer on ice for 30 minutes. For intracellular cytokine staining, cells were fixed and permeabilized in buffer and stained with antibodies to detect intracellular cytokines or transcription factors. All samples were acquired on LSRII flow cytometers and analyzed with Flowjo (Flowjo, LLC., Ashland, Or). Antibodies against CD45 (A20), CD8 (53-6.7) and CD3 (145-2C11) were purchased from eBioscience. The antibodies against CD4 (RM4-5), PD-1 (RMP1-14), and TIGIT (1G9) were purchased from Biolegend.
Wang J., Perry C.J., Meeth K., Thakral D., Damsky W., Micevic G., Kaech S., Blenman K, & Bosenberg M. (2017). UV-induced somatic mutations elicit a functional T cell response in the YUMMER1.7 Mouse Melanoma Model. Pigment cell & melanoma research, 30(4), 428-435.
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5

Multi-lineage Hematopoietic Engraftment Analysis

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Peripheral blood was stained with lineage-specific antibodies and analyzed on LSRII (BD Biosciences) to assess engraftment. The following antibodies were used: Gr1 (RB6-8C5), Mac1 (M1/70), B220 (RA3-B2), CD3 (145-2C11), and Ter119 from eBioscience; CD45.1 and CD45.2 from BD Biosciences. The CD45.1 chimerisms in non-irradiated, untransplanted CD45.2 mice were used as a negative staining control. Recipients with multi-lineage chimerism above the average negative-control chimerism plus 3 standard deviations were considered to have multi-lineage engraftment (Fig. 4f).
Li P., Lahvic J.L., Binder V., Pugach E.K., Riley E.B., Tamplin O.J., Panigrahy D., Bowman T.V., Barrett F.G., Heffner G.C., McKinney-Freeman S., Schlaeger T.M., Daley G.Q., Zeldin D.C, & Zon L.I. (2015). Epoxyeicosatrienoic Acids Enhance Embryonic Haematopoiesis and Adult Marrow Engraftment. Nature, 523(7561), 468-471.
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6

Multi-lineage Hematopoietic Engraftment Analysis

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Peripheral blood was stained with lineage-specific antibodies and analyzed on LSRII (BD Biosciences) to assess engraftment. The following antibodies were used: Gr1 (RB6-8C5), Mac1 (M1/70), B220 (RA3-B2), CD3 (145-2C11), and Ter119 from eBioscience; CD45.1 and CD45.2 from BD Biosciences. The CD45.1 chimerisms in non-irradiated, untransplanted CD45.2 mice were used as a negative staining control. Recipients with multi-lineage chimerism above the average negative-control chimerism plus 3 standard deviations were considered to have multi-lineage engraftment (Fig. 4f).
Li P., Lahvic J.L., Binder V., Pugach E.K., Riley E.B., Tamplin O.J., Panigrahy D., Bowman T.V., Barrett F.G., Heffner G.C., McKinney-Freeman S., Schlaeger T.M., Daley G.Q., Zeldin D.C, & Zon L.I. (2015). Epoxyeicosatrienoic Acids Enhance Embryonic Haematopoiesis and Adult Marrow Engraftment. Nature, 523(7561), 468-471.
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7

Comprehensive Immune Profiling by Flow Cytometry

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Flow cytometry was performed with Life Attune Nxt FACS machine (Life Technology/Thermo Fisher Scientific, Waltham, MA). Fluoro-chrome conjugated anti-mouse CD45 (30-F11, Biolegend), CD3 (145–2C11, eBioscience), CD4 (RM4-5, eBioscience), CD8 (eBioH35-17.2, eBioscience), Foxp3 (FJK-16s, eBioscience), Ki-67 (SolA15, eBioscience), Granzyme B (NGZB, eBioscience), Ly-6C (AL-21, Biolegend), Ly-6G (1A8, Biolegend), mPD-L1 (10 F.9G2, Biolegend), hPD-L1 (29E.29A3, Biolegend) and IFN-gamma (XMG1.2). Granzyme B and IFN-gamma staining was performed with intracellular staining kit from BD Bioscience. While Ki-67 and Foxp3 staining was done with nuclear protein staining kit from eBioscience (San Diego, CA). FACS data were analyzed using FlowJo software (Tree Star, Ashland, OR).
Huang A., Peng D., Guo H., Ben Y., Zuo X., Wu F., Yang X., Teng F., Li Z., Qian X, & Qin F.X. (2017). A human programmed death-ligand 1-expressing mouse tumor model for evaluating the therapeutic efficacy of anti-human PD-L1 antibodies. Scientific Reports, 7, 42687.
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8

Multiparametric Flow Cytometry Analysis

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For flow cytometry staining, dissociated cells were incubated with a fixable viability stain 510 (BD, Biosciences) followed by staining with primary antibodies directed against CD45 (30-F11), CD11b (M1/70), Gr1 (RB6-8C5), F4/80 (BM8), CD206 (C068C2), MHCII (AF6-120.1), CD44 (IM7), PDL1 (MIH5), TIGIT (1G9), CD69 (H1.2F3) obtained from BD Biosciences; CD3 (145-2C11), CD4 (GK1.5), CD8 (53–6.7), pSykY348 (moch1ct) from eBioscience, PD1 (29F.1A12), and CD62L (MEL-14) from Biolegend. Samples were acquired on BD Fortessa or LSRII and data were analyzed using FlowJo software version 10 (Treestar, Ashland, OR, USA).
Rohila D., Park I.H., Pham T.V., Weitz J., Hurtado de Mendoza T., Madheswaran S., Ishfaq M., Beaman C., Tapia E., Sun S., Patel J., Tamayo P., Lowy A.M, & Joshi S. (2023). Syk Inhibition Reprograms Tumor-Associated Macrophages and Overcomes Gemcitabine-Induced Immunosuppression in Pancreatic Ductal Adenocarcinoma. Cancer Research, 83(16), 2675-2689.
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9

Comprehensive Immune Cell Profiling

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Cellular phenotypic analysis was carried out by direct immunofluorescence with the following antibodies: CD11b M1/70 (eBioscience), Ly6g 1A8 (eBioscience) F4/80 BM8 (Biolegend), MHCII 2G9 (Biosciences), NK1.1 S17016D (Biolegend), CD3 145-2C11 (eBioscience), CD31 390 (Biolegend), αSMA (α smooth muscle actin) 1A4 (eBioscience), CD62-E REA369 (Miltenyi Biotec), CD62P RMP-1 (Biolegend), CD54 (Biolegend), CD106 (Biolegend), CD192 (CCR2) REA538 (Miltenyi Biotec)., PSGL-1 34 RA10 (Thermofisher), CD44 IM7.8.1 (Miltenyi Biotec), CD11a 121.7 (Thermofisher), CD49d 9C10 MFR4.B (Biolegend). All incubations were done in the presence of 50 μg/ml mouse IgG or human IgG. The same isotype control antibody was always included as a negative control, and dead cells were excluded by Annexin V staining (Sigma, USA). Flow cytometry was performed with a 10 color Gallios device (Beckman Coulter, USA), cells were counted using Flow-Count fluorospheres (Beckman Coulter, USA), following the manufacturer’s instructions, and data were analyzed using Flowjo software (Tree Star, Inc, USA).
Latorre M.C., Gómez‐Oro C., Olivera‐Valle I., Blazquez‐Lopez E., Gallego‐Valle J., Ibañez‐Escribano A., Casesnoves P., González‐Cucharero C., Muñoz‐Fernandez M.A., Sanz L., Vaquero J., Martín‐Rabadań P., Perez‐Milan F, & Relloso M. (2022). Vaginal neutrophil infiltration is contingent on ovarian cycle phase and independent of pathogen infection. Frontiers in Immunology, 13, 1031941.
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10

T. muris Immune Response Analysis

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Mesenteric lymph node cells from T. muris-treated mice were isolated and single-cell suspensions were plated at 4 × 106 per ml in the medium or in the presence of antibodies against CD3 (145-2C11) and CD28 (37.51); 1 μg ml−1 each; (eBioscience, San Diego, CA) or T. muris antigen (50 μg ml-1) for 72 h. Cytokine production from cell-free supernatants was determined by standard sandwich enzyme-linked immunosorbent assay (ELISA) using commercially available antibodies (eBioscience). T. muris-specific serum IgG1 and IgG2a levels were determined by ELISA on plates coated with T. muris Ag (5 μg ml-1). Total protein was isolated from fecal samples, resolved by sodium dodecyl sulfate- polyacrylamide gel electrophoresis, and immunoblotted using a rabbit anti-mouse RELM-β antibody (PeproTech, Rocky Hill, NJ).
Oudhoff M.J., Antignano F., Chenery A.L., Burrows K., Redpath S.A., Braam M.J., Perona-Wright G, & Zaph C. (2016). Intestinal Epithelial Cell-Intrinsic Deletion of Setd7 Identifies Role for Developmental Pathways in Immunity to Helminth Infection. PLoS Pathogens, 12(9), e1005876.
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