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Anti cd3ε 145 2c11

Manufactured by BioXCell
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Anti-CD3ε (145-2C11) is a monoclonal antibody that recognizes the CD3ε chain of the T cell receptor complex. It is commonly used in research applications to activate and stimulate T cells.

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Lab products found in correlation

Anti cd28 37, by BioXCell (3 mentions) Rbc lysing buffer, by BioLegend (1 mentions) 40 m nylon filter, by BD (1 mentions) Anti cd28 antibodies, by BioXCell (1 mentions) Rpmi 1640 medium, by Corning (1 mentions) Facsaria 3 cell sorter, by BD (1 mentions) Monensin, by Thermo Fisher Scientific (1 mentions) Facsaria 3, by BD (1 mentions) Recombinant mouse il 6, by Thermo Fisher Scientific (1 mentions) Recombinant human tgf β1, by Thermo Fisher Scientific (1 mentions) Cd4 t cell isolation kit, by Miltenyi Biotec (1 mentions) Brefeldin a, by Thermo Fisher Scientific (1 mentions) Ionomycin, by Merck Group (1 mentions) Phorbol myristate acetate (pma), by Merck Group (1 mentions) 96 well flat bottom plate, by Corning (1 mentions) C57bl 6j mice, by Charles River Laboratories (1 mentions) 2 nbdg, by Cayman Chemical (1 mentions)

Close spelling variants of this product

Anti-CD3ε (clone 145-2C11) Anti-CD3ε mAb (clone 145-2C11, Monoclonal anti-mouse CD3ε [145–2c11] Anti-CD3ε (2C11; Anti-CD3ε

5 protocols using anti cd3ε 145 2c11

1

Murine T Cell Isolation and Culture

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The spleens of 6‐to 8‐week‐old C1QBP+/− and C1QBP+/+ C57/BL6 mice were harvested, gently ground in MACS buffer (PBS + 0.5% BSA + 2 mM EDTA) and then passed through a sterile 40‐µm nylon filter (BD Falcon). Red blood cells were lysed with RBC Lysing Buffer (BioLegend). The murine T cells were isolated by Pan T magnetic Microbeads (Miltenyi Biotec) from splenocytes obtained from C1QBP+/+ and C1QBP+/− mice. CD4+ or CD8+ T cells were sorted on a FACSAria III Cell Sorter (BD Biosciences) and planted in 24‐well plates coated with 2.5 μg/mL anti–CD3ε (145‐2C11, Bio‐X‐Cell) and 1 μg/mL anti–CD28 antibodies (37.51 Bio‐X‐Cell). All the above cells were cultured in RPMI‐1640 medium (Corning) supplemented with 10% FBS, 4 mM l‐glutamine, 1% penicillin/streptomycin, 2 ng/mL IL‐2, 10 ng/mL IL‐7, 5 ng/mL IL‐15, and 75 μM β‐mercaptoethanol and incubated at 37°C in a 5% CO2 humidified atmosphere.
Tian H., Wang G., Wang Q., Zhang B., Jiang G., Li H., Chai D., Fang L., Wang M, & Zheng J. (2022). Complement C1q binding protein regulates T cells’ mitochondrial fitness to affect their survival, proliferation, and anti–tumor immune function. Cancer Science, 113(3), 875-890.
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2

Naïve CD4+ T Cell Polarization

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CD4+ T cells were isolated from the lymph nodes and spleen of Tgfb1fl/flIl17aCreR26YFP and WT control mice with the CD4+ T Cell Isolation Kit (Miltenyi Biotec, Bergisch Gladbach, Germany), and then naïve CD4+ T cells (CD4+CD25−CD44lowCD62Lhigh) were sorted using a BD FACSAriaTM III (BD Biosciences, San Jose, CA, USA). Anti-CD3ε (145-2C11, 1 µg/mL) (BioXCell, West Lebanon, NH, USA) and anti-CD28 (37.51, 1 µg/mL) (BioXCell) Abs were used to precoat a 96-well flat-bottom plate (Corning, Steuben Country, NY, USA) overnight at 4 °C. After washing the plate with PBS, naïve CD4+ T cells (1 × 105 cells/well) were cultured with recombinant mouse IL-6 (10 ng/mL) (PeproTech, Rocky Hill, NJ, USA) and recombinant human TGF-β1 (1 and 5 ng/mL) (PeproTech) for 72–96 h. The cultured cells were treated with 100 ng/mL PMA (Sigma-Aldrich, Saint Louis, MO, USA), 1 µM ionomycin (Sigma-Aldrich), brefeldin A (Thermo Fisher Scientific, Waltham, MA, USA), and monensin (Thermo Fisher Scientific) for an additional 3–6 h before flow cytometric analysis.
Choi G., Park Y.J., Cho M., Moon H., Kim D., Kang C.Y., Chung Y, & Kim B.S. (2021). A critical role for Th17 cell-derived TGF-β1 in regulating the stability and pathogenicity of autoimmune Th17 cells. Experimental & Molecular Medicine, 53(5), 993-1004.
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3

Single-cell RNA-seq of Activated OT-I CD8+ T Cells

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Naïve WT (CD45.1+) or PSGL-1−/− (CD90.1+) OT-I CD8+ T cells were activated with immobilized anti-CD3ε (145–2C11, BioXcell) and anti-CD28 (37.51, BioXcell)(5 μg/mL, each) for 3 days prior to transfer. 1×106 activated WT or PSGL-1−/− OT-I CD8+ T cells were then injected I.V. into tumor-bearing C57BL/6 (CD45.2+/CD90.2+) mice, 7 days after inoculation with B16-OVA tumors. Six days after T cell transfer, live donor OT-I CD8+ T cells were sorted by FACS from B16-OVA tumors. Live sorted cells were immediately processed on a 10X Chromium (10X) to generate single cell 3’ libraries using the 10X Single Cell 3’ Reagent Kits v2 as per (manufacturer’s instructions (Rev E, 2018). Data were acquired using a HiSeq 400 PE50 at the Institute for Genomic Medicine Facility at the University of California, San Diego.
Hope J.L., Otero D.C., Bae E.A., Stairiker C.J., Palete A.B., Faso H.A., Lin M., Henriquez M.L., Roy S., Seo H., Lei X., Wang E.S., Chow S., Tinoco R., Daniels G.A., Yip K., Campos A.R., Yin J., Adams P.D., Rao A, & Bradley L.M. (2023). PSGL-1 attenuates early TCR signaling to suppress CD8+ T cell progenitor differentiation and elicit terminal CD8+ T cell exhaustion. Cell reports, 42(5), 112436.
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4

Single-cell RNA-seq of Activated OT-I CD8+ T Cells

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Naïve WT (CD45.1+) or PSGL-1−/− (CD90.1+) OT-I CD8+ T cells were activated with immobilized anti-CD3ε (145–2C11, BioXcell) and anti-CD28 (37.51, BioXcell)(5 μg/mL, each) for 3 days prior to transfer. 1×106 activated WT or PSGL-1−/− OT-I CD8+ T cells were then injected I.V. into tumor-bearing C57BL/6 (CD45.2+/CD90.2+) mice, 7 days after inoculation with B16-OVA tumors. Six days after T cell transfer, live donor OT-I CD8+ T cells were sorted by FACS from B16-OVA tumors. Live sorted cells were immediately processed on a 10X Chromium (10X) to generate single cell 3’ libraries using the 10X Single Cell 3’ Reagent Kits v2 as per (manufacturer’s instructions (Rev E, 2018). Data were acquired using a HiSeq 400 PE50 at the Institute for Genomic Medicine Facility at the University of California, San Diego.
Hope J.L., Otero D.C., Bae E.A., Stairiker C.J., Palete A.B., Faso H.A., Lin M., Henriquez M.L., Roy S., Seo H., Lei X., Wang E.S., Chow S., Tinoco R., Daniels G.A., Yip K., Campos A.R., Yin J., Adams P.D., Rao A, & Bradley L.M. (2023). PSGL-1 attenuates early TCR signaling to suppress CD8+ T cell progenitor differentiation and elicit terminal CD8+ T cell exhaustion. Cell reports, 42(5), 112436.
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5

Metabolic Profiling in C57BL/6J Mice

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C57BL/6J mice were purchased from Charles River. Mice were bred and maintained at the Instituto de Medicina Molecular animal facilities, Lisbon. Mice were systematically compared with cohoused littermate controls unless stated otherwise; 8- to 14-wk-old male and female mice, age and sex matched, were used. All mice were housed in individually ventilated cages (IVC) with temperature-controlled conditions under a 12-h light/dark cycle, and the mice were kept in specific-pathogen-free conditions with free access to drinking water and food. Anti-CD3ε (145-2C11, BioXCell) was given intraperitoneally (i.p.) at 25 μg per animal, 2-NBDG (CAY11046, Cayman Chemical) was given with a retroorbital injection of 100 or 300 µg 1 h before killing. Glucose 10% wt/vol was provided in the drinking water. All animal experimentation complied with the European Union directive and regulations of the Direção-Geral de Alimentação e Veterinária Portugal and local ethical review committee and guidelines.
Konjar Š., Ferreira C., Carvalho F.S., Figueiredo-Campos P., Fanczal J., Ribeiro S., Morais V.A, & Veldhoen M. (2022). Intestinal tissue-resident T cell activation depends on metabolite availability. Proceedings of the National Academy of Sciences of the United States of America, 119(34), e2202144119.
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