Naïve CD4+ T cells were forced to polarize into Th17 cells by 3 days of culture under Th17-differentiating conditions using a protocol and reagents provided by BioLegend (unless otherwise indicated). Briefly, isolated naïve CD4+ T cells were suspended in ‘full medium’, consisting of RPMI 1640 medium (ThermoFisher), supplemented with 10% FBS (Biochrom) and 1% penicillin/streptomycin (ThermoFisher), and added to multiwell plates precoated with anti-mouse CD3 mAb (clone 145-2C11, 5 µg/ml). To induce Th17 differentiation, recombinant IL-6 (50 ng/ml), recombinant human TGF-β (1 ng/ml), recombinant mouse IL-23 (5 ng/ml), anti-mouse IL-4 mAb (clone 11B11, 10 µg/ml), anti-mouse IFN-γ mAb (clone XMG1.2, 10 µg/ml) and anti-mouse CD28 mAb (clone 37.51, 5 µg/ml, BD Biosciences) were added to the cultures.
Anti mouse cd28 mab clone 37
Manufactured by BD
Sourced in United States
Anti-mouse CD28 mAb (clone 37.51) is a monoclonal antibody that binds to the mouse CD28 receptor. CD28 is a co-stimulatory molecule expressed on the surface of T cells that plays a crucial role in T cell activation and proliferation. This antibody can be used in various immunological applications to study T cell biology.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Harvard Bioscience (1 mentions)
Mouse il 15, by BioLegend (1 mentions)
Fetal calf serum (fcs), by BioLegend (1 mentions)
Rpmi 1640 medium, by BioLegend (1 mentions)
2 protocols using anti mouse cd28 mab clone 37
1
Th17 Cell Differentiation Protocol
2
Activation and Retroviral Modification of CAR T Cells
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CAR T cells were activated in vitro by murine IL-2 (100 U/mL) (ImmunoTools, Friesoythe, Germany), the anti-mouse CD3e monoclonal antibody (mAb) clone 145-2C11 (200 ng/mL), and the anti-mouse CD28 mAb clone 37.51 (100 ng/mL) (BD Biosciences, Mountain View, CA, USA). T cells were cultivated in RPMI 1640 medium (Dutch modification), 10% (v/v) FCS, mouse IL-15 (5 ng/mL) (BioLegend, San Diego, CA, USA), 1% (v/v) non-essential amino acids, and 2% (v/v) L-glutamate. To produce the retroviruses, 293T cells were co-transfected with the retroviral Moloney murine leukemia virus (MMLV)-based helper plasmids coding for the gag, pol, and env genes together with the CAR, IL-12, or IL-18 expression construct, respectively. The retroviral modification of mouse T cells was previously described in detail (John et al., 2014) .
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