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Anti cd3ε 2c11

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Anti-CD3ε (2C11) is a monoclonal antibody that binds to the CD3ε subunit of the T cell receptor (TCR) complex. It is commonly used in research applications to activate and stimulate T cells in vitro.

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Lab products found in correlation

Anti cd28 37, by BioXCell (2 mentions) Interleukin 2 (il 2), by Thermo Fisher Scientific (2 mentions) Magnetic bead negative selection, by Miltenyi Biotec (1 mentions) Efluor670 proliferation dye, by Thermo Fisher Scientific (1 mentions) Anti igm, by Jackson ImmunoResearch (1 mentions) Facsaria 2, by BD (1 mentions) Anti ifn γ xmg1, by BioXCell (1 mentions) Facsariatm 3, by BD (1 mentions) Cd4 t cell isolation kit, by Miltenyi Biotec (1 mentions) Tgf β1, by Thermo Fisher Scientific (1 mentions) 96 well flat bottom plate, by Corning (1 mentions)

3 protocols using anti cd3ε 2c11

1

Retroviral Transduction of SMARTA CD4+ T Cells

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The oligonucleotides incorporating miR-30 microRNA sequence into target sequences (Table S1) were cloned into the plasmid pLMPd-Ametrine. cDNA encoding GAPDH or ALDOA was subcloned into pMIG-myc-GFP vector. Plat-E packaging cells were transfected with 2.5 µg of retroviral vector with 7.5 µl of TransIT-LT1 transfection reagent (Mirus). The culture supernatant containing retrovirus was collected at 48 h after transfection. Naive SMARTA CD4+ T cells were stimulated with plate-bound anti-CD3ε (2C11; BioXCell) and anti-CD28 (37.51; BioXCell) for 24 h, then infected with retrovirus together with 8 µg/ml polybrene and 40 ng/ml IL-2 (Peprotech) by centrifugation at 1,900 rpm for 90 min at 37°C. At 24 h after infection, the cells were rested in fresh medium with 40 ng/ml IL-2 for 48 h followed by culturing in fresh medium in the presence of 2 ng/ml IL-7 for 12–16 h.
Zhu Y., Zhao Y., Zou L., Zhang D., Aki D, & Liu Y.C. (2019). The E3 ligase VHL promotes follicular helper T cell differentiation via glycolytic-epigenetic control. The Journal of Experimental Medicine, 216(7), 1664-1681.
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2

Flow Sorting and Co-Culture of T and B Cells

Cited 2 times
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T and B cells from allograft-draining lymph nodes (DLN) were enriched, flow sorted and co-cultured as previously described (31 (link)). Briefly, magnetic bead negative selection (Miltenyi Biotec) was used to enrich CD4+ T and CD19+ B cells. T cells from DLNs were flow sorted into Tfh (CD19−CD4+PD-1hiCXCR5+GITR−) and Tfr (CD19−CD4+PD-1hiCXCR5+GITR+) cells on a FACSAria II (BD Biosciences). For proliferation assessment, B cells were stained with eFluor670 proliferation dye (eBioscience). 3×104 Tfh cells were cultured with 5×104 B cells with or without 1.5×104 Tfr cells in 96-well plates in anti-CD3ε (2C11, 2 μg/mL, BioXcell) and anti-IgM (5 μg/mL, Jackson Immunoresearch) containing media for 5 days. Culture media was supplemented with immunosuppression where indicated at the following concentrations: CTLA-4-Ig (50 μg/mL), anti-CD28 dAb (25 μg/mL), anti-CTLA-4 mAb (50 μg/mL). Anti-CD28 dAb and CTLA-4-Ig dosing was based on molecular weight, serum half-life, and murine mixed lymphocyte reaction EC50 (14 (link), 20 (link)).
La Muraglia GM I.I., Zeng S., Crichton E.S., Wagener M.E., Ford M.L, & Badell I.R. (2020). Superior inhibition of alloantibody responses with selective CD28 blockade is CTLA-4-dependent and T follicular helper cell-specific. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons, 21(1), 73-86.
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3

Treg Cell Differentiation from Naïve CD4 T Cells

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CD4 T cells were isolated from lymph nodes and spleens of Foxp3YFP-CreTgfb1fl/+ or Foxp3YFP-CreTgfb1fl/fl mice with a CD4+ T Cell Isolation Kit (Miltenyi Biotec, Bergisch Gladbach, Germany), and then naïve CD4 T cells (CD4+CD25−CD44lowCD62Lhigh) were sorted using FACSAriaTM III (BD Biosciences, San Jose, CA, USA). For Treg cell differentiation, anti-CD3ε (2C11, 1 µg/mL) (BioXcell, West Lebanon, NH, USA) and anti-CD28 (37.51, 1 µg/mL) (BioXcell, West Lebanon, NH, USA) Abs were pre-coated in a 96-well flat bottom plate (Corning, Steuben Country, NY, USA) overnight at 4 °C. After washing the plate with phosphate-buffered saline (GenDEPOT, Katy, TX, USA), naïve CD4 T cells (1 × 105 cells/well) were cultured with IL-2 (20 ng/mL) (PeproTech, Rocky Hill, NJ, USA), TGF-β1 (3 ng/mL) (PeproTech, Rocky Hill, NJ, USA), and anti-IFN-γ (XMG1.2, 1 µg/mL) (BioXcell, West Lebanon, NH, USA) for 96 h.
Choi G., Na H., Kuen D.S., Kim B.S, & Chung Y. (2020). Autocrine TGF-β1 Maintains the Stability of Foxp3+ Regulatory T Cells via IL-12Rβ2 Downregulation. Biomolecules, 10(6), 819.
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