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Anti cd3 145 2c11

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Anti-CD3 (145-2C11) is a monoclonal antibody that recognizes the CD3 complex on T cells. It is commonly used in research applications to activate and stimulate T cell responses.

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Lab products found in correlation

Celltrace violet, by Thermo Fisher Scientific (2 mentions) Dynabead, by Thermo Fisher Scientific (2 mentions) Iscove modified dulbecco media (imdm), by Merck Group (2 mentions) Anti hamster antibody, by MP Biomedicals (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Sodium pyruvate, by Thermo Fisher Scientific (2 mentions) Recombinant mouse il 2, by Thermo Fisher Scientific (2 mentions) L glutamine, by Thermo Fisher Scientific (2 mentions) Hepes, by Thermo Fisher Scientific (2 mentions) Biotinylated antibodies, by BioLegend (2 mentions) Anti cd3 145 2c11, by BioLegend (2 mentions) Anti cd28 37, by BioLegend (2 mentions) β mercaptoethanol, by Thermo Fisher Scientific (2 mentions) Magnetic microbead selection, by Miltenyi Biotec (2 mentions) Human tgf β1, by BioLegend (2 mentions) Click s media, by Merck Group (2 mentions)

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Anti-CD3 (24 citations)

4 protocols using anti cd3 145 2c11

1

Treg Differentiation and Suppression Assay

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Unless otherwise noted, Tregs were cultured in RPMI-1640 medium (plus β-mercaptoethanol) supplemented with 10% fetal bovine serum, 1% L-glutamine, 1% penicillin-streptomycin, 50 ng/ml IL-2 and anti-CD3/CD28 coated Dynabeads (Thermo Fisher) at 1:1 ratio (cells: beads). For glucose challenge experiments, glucose-free RPMI-1640 medium (Thermo Fisher, 1187020) were used. 2-DG was purchased from Sigma (D8375). For in vitro iTreg differentiation, 24-well plates were pre-coated with 40 μg/ml anti-hamster antibody (MP Biomedical). CD4+CD25−CD44−CD62L+ naive T cells were sorted and cultured for 3 days in IMDM (Sigma-Aldrich) supplemented with 0.25 μg/ml anti-CD3 (145-2C11), 1 μg/ml anti-CD28 (37.51), 2 μg/ml anti-IL-4 (11B11), 2 αg/ml anti-IFN-γ (XMG1.2) and 5 ng/ml TGF-β. For in vitro Treg suppression assay, splenocytes from CD45.1+ mice were labeled with Cell Trace Violet (Thermo Fisher) and cultured with or without various ratio of sorted splenic Tregs from control or Tfamfl/flFoxp3Yfp-Cre mice. Cells were cultured for 3d with soluble 1 μg/ml anti-CD3/CD28. The division of CD45.1+CD4+ T cells were assessed by the dilution of Cell Trace Violet.
Fu Z., Ye J., Dean J.W., Bostick J.W., Weinberg S.E., Xiong L., Oliff K.N., Chen Z.E., Avram D., Chandel N.S, & Zhou L. (2019). Requirement of Mitochondrial Transcription Factor A in Tissue-Resident Regulatory T Cell Maintenance and Function. Cell reports, 28(1), 159-171.e4.
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2

Treg Differentiation and Suppression Assay

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Unless otherwise noted, Tregs were cultured in RPMI-1640 medium (plus β-mercaptoethanol) supplemented with 10% fetal bovine serum, 1% L-glutamine, 1% penicillin-streptomycin, 50 ng/ml IL-2 and anti-CD3/CD28 coated Dynabeads (Thermo Fisher) at 1:1 ratio (cells: beads). For glucose challenge experiments, glucose-free RPMI-1640 medium (Thermo Fisher, 1187020) were used. 2-DG was purchased from Sigma (D8375). For in vitro iTreg differentiation, 24-well plates were pre-coated with 40 μg/ml anti-hamster antibody (MP Biomedical). CD4+CD25−CD44−CD62L+ naive T cells were sorted and cultured for 3 days in IMDM (Sigma-Aldrich) supplemented with 0.25 μg/ml anti-CD3 (145-2C11), 1 μg/ml anti-CD28 (37.51), 2 μg/ml anti-IL-4 (11B11), 2 αg/ml anti-IFN-γ (XMG1.2) and 5 ng/ml TGF-β. For in vitro Treg suppression assay, splenocytes from CD45.1+ mice were labeled with Cell Trace Violet (Thermo Fisher) and cultured with or without various ratio of sorted splenic Tregs from control or Tfamfl/flFoxp3Yfp-Cre mice. Cells were cultured for 3d with soluble 1 μg/ml anti-CD3/CD28. The division of CD45.1+CD4+ T cells were assessed by the dilution of Cell Trace Violet.
Fu Z., Ye J., Dean J.W., Bostick J.W., Weinberg S.E., Xiong L., Oliff K.N., Chen Z.E., Avram D., Chandel N.S, & Zhou L. (2019). Requirement of Mitochondrial Transcription Factor A in Tissue-Resident Regulatory T Cell Maintenance and Function. Cell reports, 28(1), 159-171.e4.
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3

In Vitro Treg Cell Induction

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Treg cell induction in vitro was performed by culturing 0.2 × 106 sorted T cells/well in 96 well plates pre-coated with anti-CD3 (145–2C11) (1 μg/mL) using Click’s media (Millipore-Sigma) containing 10% fetal bovine serum (FBS) (Gemini); penicillin-streptomycin, L-glutamine, HEPES, β-mercaptoethanol, sodium pyruvate (all from GIBCO); recombinant mouse IL-2 (200 units/ml), human TGF-β1 (0.25 ng/ml, or as indicated in figure legends of individual figures), and anti-CD28 (37.51) (1.5 μg/mL) (Biolegend).
Th0 cultures were prepared by culturing 0.2 × 106 sorted T cells/well with 0.1 × 106 antigen presenting cells (APCs) in the presence of anti-CD3 (145–2C11) (1 μg/ml) (Biolegend) using Click’s media (Millipore-Sigma) containing 10% FBS (Gemini) and penicillin-streptomycin, L-glutamine, β-mercaptoethanol, HEPES, and sodium pyruvate (all from GIBCO) for 1–3 days. The APCs used in these cultures were isolated from pooled lymph node and spleen cells depleted of T cells (CD4+, CD8+, CD3+) and CD49b+ NK cells using biotinylated antibodies (Biolegend) and magnetic microbead selection (Miltenyi).
Opejin A., Surnov A., Misulovin Z., Pherson M., Gross C., Iberg C.A., Fallahee I., Bourque J., Dorsett D, & Hawiger D. (2020). A Two-Step Process of Effector Programming Governs CD4+ T Cell Fate Determination Induced by Antigenic Activation in the Steady State. Cell reports, 33(8), 108424.
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4

In Vitro Treg Cell Induction

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Treg cell induction in vitro was performed by culturing 0.2 × 106 sorted T cells/well in 96 well plates pre-coated with anti-CD3 (145–2C11) (1 μg/mL) using Click’s media (Millipore-Sigma) containing 10% fetal bovine serum (FBS) (Gemini); penicillin-streptomycin, L-glutamine, HEPES, β-mercaptoethanol, sodium pyruvate (all from GIBCO); recombinant mouse IL-2 (200 units/ml), human TGF-β1 (0.25 ng/ml, or as indicated in figure legends of individual figures), and anti-CD28 (37.51) (1.5 μg/mL) (Biolegend).
Th0 cultures were prepared by culturing 0.2 × 106 sorted T cells/well with 0.1 × 106 antigen presenting cells (APCs) in the presence of anti-CD3 (145–2C11) (1 μg/ml) (Biolegend) using Click’s media (Millipore-Sigma) containing 10% FBS (Gemini) and penicillin-streptomycin, L-glutamine, β-mercaptoethanol, HEPES, and sodium pyruvate (all from GIBCO) for 1–3 days. The APCs used in these cultures were isolated from pooled lymph node and spleen cells depleted of T cells (CD4+, CD8+, CD3+) and CD49b+ NK cells using biotinylated antibodies (Biolegend) and magnetic microbead selection (Miltenyi).
Opejin A., Surnov A., Misulovin Z., Pherson M., Gross C., Iberg C.A., Fallahee I., Bourque J., Dorsett D, & Hawiger D. (2020). A Two-Step Process of Effector Programming Governs CD4+ T Cell Fate Determination Induced by Antigenic Activation in the Steady State. Cell reports, 33(8), 108424.
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