CD8 T cells were purified from spleens by negative selection using EasySep™ Mouse CD8+ T Cell Isolation Kit (Stem Cell) according to manufacturer’s instructions. Cells were stimulated with 100 U/ml recombinant human IL-2, 1 μg/ml anti-mouse CD3ε (145-2C11, Biolegend) and 5 μg/ml anti-mouse CD28 (37.51, eBiosciences) in RPMI-1640 medium with 10% FBS, 10 mM HEPES (Gibco), 100 μM NEAA (Gibco), 50 U/ml penicillin, 50 μg/ml streptomycin and 50 μM β-mercaptoethanol.
Anti mouse cd3ε 145 2c11
Manufactured by BioLegend
Anti-mouse CD3ε (145-2C11) is a monoclonal antibody that binds to the CD3ε chain of the T cell receptor complex in mice. This antibody can be used for the identification and characterization of T cells in flow cytometry and other immunological applications.
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Lab products found in correlation
Anti mouse cd28 37, by Thermo Fisher Scientific (1 mentions)
Easysep mouse cd8 t cell isolation kit, by STEMCELL (1 mentions)
Hepes, by Thermo Fisher Scientific (1 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions)
β actin 8h10d10, by Cell Signaling Technology (1 mentions)
Apotracker green, by BioLegend (1 mentions)
Mouse il 4, by BioLegend (1 mentions)
Biotin anti mouse lineage panel, by BioLegend (1 mentions)
Mojosort mouse cd8 t cell isolation kit, by BioLegend (1 mentions)
Mouse il 33, by BioLegend (1 mentions)
Mouse il 2, by BioLegend (1 mentions)
2 protocols using anti mouse cd3ε 145 2c11
1
Isolation and Stimulation of CD8 T Cells
2
Isolation and Culture of Mouse T Cells
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Mouse IL-2 (catalog 575402), mouse IL-4 (catalog 547302), mouse IL-33 (catalog 580502), anti-mouse CD3ε (145-2C11), anti-mouse CD28 (37.51), human TGF-β1 (catalog 781802), anti-mouse IL-4 mAb (clone 11B11), anti-mouse IFN-γ mAb (XMG1.2), and Apotracker Green were purchased from BioLegend. Antibodies for TRAF4 (D1N3A), MyD88 (D80F5), p-mTOR (S2448, D9E), mTOR (7C10), p-AKT (S473, D9E), p-AKT (T308, D25E6), AKT (11E7), p-JNK (T183/Y185, 81E11), SAPK/JNK (no. 9252), p-p38 (T180/Y182, D3F9), p38 (D13E1), p-ERK1/2 (T202/Y204, D13.14.4E), Erk1/2 (137F5), p-IκBα (S32, 14D4), IκBα (L35A5), TRAF6 (D21G3), and β-Actin (8H10D10) were obtained from Cell Signaling Technology. Mouse T1/ST2 antibody (DJ8) and rabbit polyclonal against ST2 (ab228543) were obtained from MD Bioproducts and Abcam, respectively. AKT inhibitor VIII (CAS 612847-09-3) and LY3214996 (CAS 1951483-29-6) were purchased from Cayman Chemical. Primary mouse CD4+ T cells were cultured in complete TexMACS Medium (Miltenyi Biotec, 130-097-196) supplemented with 10% FBS, 50 μM β-mercaptoethanol, 100 U/mL penicillin, and 100 μg/mL streptomycin. CD8+ cells and ILCs were isolated from spleen using MojoSort Mouse CD8 T Cell Isolation Kit and Biotin anti-mouse Lineage Panel from BioLegend.
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