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Cd3ε 145 2c11

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The CD3ε (145–2C11) is a monoclonal antibody that recognizes the epsilon subunit of the CD3 complex, which is a critical component of the T cell receptor (TCR) complex. The CD3 complex plays a key role in T cell activation and signal transduction.

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Lab products found in correlation

Gr 1 rb6 8c5, by Thermo Fisher Scientific (2 mentions) Cd117 2b8, by BD (2 mentions) F4 80 bm8, by Thermo Fisher Scientific (2 mentions) Fcεriα mar 1, by Thermo Fisher Scientific (2 mentions) Cd49b dx5, by BioLegend (2 mentions) Ccr3 tg14, by BioLegend (2 mentions) Cd16 32 2.4g2, by BioXCell (2 mentions) Propidium iodide, by BD (2 mentions) B220 ra3 6b2, by BD (2 mentions) Nk1.1 pk136, by Thermo Fisher Scientific (2 mentions) B220 ra3 6b2, by BioLegend (1 mentions) Tc11 18h10, by BioLegend (1 mentions) Nkp46 29a1.4, by BioLegend (1 mentions) Cd4 gk1, by BioLegend (1 mentions) I a 1 e m5 114, by BioLegend (1 mentions) Cd8α 53 6, by Thermo Fisher Scientific (1 mentions) Tcrβ h57 597, by BioLegend (1 mentions) Cd25 pc61, by BioLegend (1 mentions) Ionomycin, by Merck Group (1 mentions) Phorbol myristate acetate (pma), by Merck Group (1 mentions)

Spelling variants of this product

Anti-CD3ε antibody (clone 145–2C11, (4 citations) Anti-CD3ε (clone 145-2C11; (3 citations) Anti-CD3ε (145-2C11) (2 citations) CD3ε (clone 145-2C11; (2 citations)

5 protocols using cd3ε 145 2c11

1

Comprehensive Immune Cell Profiling

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Red blood cell indices were measured by an automated hematology analyzer. Leukocyte populations in blood were assessed by flow cytometry as follows: neutrophils Gr-1+, T cells CD3+, B cells B220+, monocytes F4/80+ Gr-1−, eosinophils CCR3+ SSChi, basophils FcεRIα+ CD49b+. Peritoneal cells were isolated by peritoneal lavage with PBS. MCs were identified by staining peritoneal cells for FcεRIα and CD117 (c-kit). RBCs were lysed in ACK buffer (0.15 M NH4Cl, 1 mM KHCO3, 0.1mM EDTA). Cells were blocked with anti-CD16/32 for 10 min on ice, before staining with antibodies for flow cytometry for 30 min on ice. Propidium iodide (BD Biosciences, San Jose, California, USA) was used to exclude dead cells from analysis. Antibodies used were: FcεRIα (MAR-1; eBiosciences, San Diego, California, USA), CD117 (2B8; BD Biosciences), CD49b (DX5; Biolegend, San Diego, California, USA), CCR3 (TG14; Biolegend), B220 (RA3–6B2, BD Biosciences), CD3ε (145–2C11; eBiosciences), Gr-1 (RB6–8C5; eBiosciences), F4/80 (BM8; eBiosciences), CD16/32 (2.4G2, Bio X Cell, West Lebanon, New Hampshire, USA).
Hernandez J.D., Yu M., Sibilano R., Tsai M, & Galli S.J. (2019). Development of multiple features of antigen-induced asthma pathology in a new strain of mast cell deficient BALB/c-KitW-sh/W-sh mice. Laboratory investigation; a journal of technical methods and pathology, 100(4), 516-526.
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2

Isolation and Characterization of Murine Immune Cells

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Splenocytes were isolated as previously described(23 ). PBS-perfused kidneys and lungs were minced and digested with collagenase IV (100 μg/ml) for 30 minutes at 37 °C to prepare single cells.
Isolated cells were stained with the following antibodies specific for: TCRβ (H57-597, BioLegend), CD3ε (145-2C11, eBiosciences), CD4(GK1.5, BioLegend), CD8α (53-6.7, eBiosciences), B220 (RA3-6B2, BioLegend), CD25(PC61, BioLegend), I-A/I-E (M5/114.15.2, BioLegend), Nkp46 (29A1.4, BioLegend) for 30min at 4 °C. For intracellular staining, cells were stimulated with 20 ng/ml of phorbol-myristate acetate (PMA, Sigma) and 1 mg/ml of ionomycin(Sigma) for 4 hours, washed and stained with TCRβ, CD4, CD8, I-A/I-E. BD cytofix/cytoperm plus with Golgi stop staining kits (BD Biosciences) were used according to the manufacturer’s protocol. Antibodies of IFN-γ (XMG1.2, BioLegend) and IL-17A (TC11-18H10.1, BioLegend) were used for detecting intracellular cytokines.
Mizui M., Koga T., Lieberman L.A., Beltran J., Yoshida N., Johnson M.C., Tisch R, & Tsokos G.C. (2014). Interleukin-2 Protects Lupus-prone Mice from Multiple End-organ Damage by Limiting CD4−CD8− Interleukin-17-producing T-cells. Journal of immunology (Baltimore, Md. : 1950), 193(5), 2168-2177.
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3

Comprehensive Immune Cell Profiling

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Red blood cell indices were measured by an automated hematology analyzer. Leukocyte populations in blood were assessed by flow cytometry as follows: neutrophils Gr-1+, T cells CD3+, B cells B220+, monocytes F4/80+ Gr-1−, eosinophils CCR3+ SSChi, basophils FcεRIα+ CD49b+. Peritoneal cells were isolated by peritoneal lavage with PBS. MCs were identified by staining peritoneal cells for FcεRIα and CD117 (c-kit). RBCs were lysed in ACK buffer (0.15 M NH4Cl, 1 mM KHCO3, 0.1mM EDTA). Cells were blocked with anti-CD16/32 for 10 min on ice, before staining with antibodies for flow cytometry for 30 min on ice. Propidium iodide (BD Biosciences, San Jose, California, USA) was used to exclude dead cells from analysis. Antibodies used were: FcεRIα (MAR-1; eBiosciences, San Diego, California, USA), CD117 (2B8; BD Biosciences), CD49b (DX5; Biolegend, San Diego, California, USA), CCR3 (TG14; Biolegend), B220 (RA3–6B2, BD Biosciences), CD3ε (145–2C11; eBiosciences), Gr-1 (RB6–8C5; eBiosciences), F4/80 (BM8; eBiosciences), CD16/32 (2.4G2, Bio X Cell, West Lebanon, New Hampshire, USA).
Hernandez J.D., Yu M., Sibilano R., Tsai M, & Galli S.J. (2019). Development of multiple features of antigen-induced asthma pathology in a new strain of mast cell deficient BALB/c-KitW-sh/W-sh mice. Laboratory investigation; a journal of technical methods and pathology, 100(4), 516-526.
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4

Characterizing Murine NK Cell Activation

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Six well plates were coated overnight at 4°C with antibody, anti-NK1.1 (PK136), anti-NKp46 (29A1.4) or anti-NKG2D (A10) (eBioscience, San Diego, CA). After sacrifice splenocytes were harvested and red blood cells lysed using ACK lysis buffer (eBioscience, San Diego, CA). 6 × 106 cells were incubated either in an antibody coated, unstimulated or PMA and ionomycin treated well for five hours in the presence of GolgiPlug (BD Biosciences, San Diego, CA). Following incubation Fc receptors were blocked with anti-CD16/CD32 (2.4G2) antibody (BD Biosciences, San Diego, CA). Invitrogen fixable aqua live/dead stain was used according to manufacturer’s instructions for dead cell exclusion. NK1.1 (PK136) (for all stimulation conditions other than samples stimulated with anti-NK1.1 antibody) or NKp46 (for samples stimulated with anti-NK1.1 antibody) and CD3ε (145-2C11) surface markers were stained (eBioscience, San Diego, CA). Cells were fixed and permeabilized using BD Cytofix/Cytoperm™ kit (BD Biosciences, San Diego, CA) according to manufactures instructions. Fc receptors were blocked with anti-CD16/CD32 (2.4G2) antibody. Intracellular staining was performed using an anti-IFNγ (XMG1.2) antibody (eBioscience, San Diego, CA). Cells were analyzed using a BD LSRFortessa cell analyzer (BD Biosciences, San Diego, CA)
Gumbleton M., Vivier E, & Kerr W.G. (2015). SHIP1 Intrinsically Regulates NK Cell Signaling and Education Resulting in Tolerance of a MHC-I Mismatched BM Graft in Mice. Journal of immunology (Baltimore, Md. : 1950), 194(6), 2847-2854.
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5

Comprehensive Immune Cell Profiling

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The following anti-mouse antibodies were used for surface staining: CD3ε (145-2C11, eBioscience), CD11b (M1/70, eBioscience), CD11c (HL3, BD), CD16/32 (101302, Biolegend), CD45 (30-F11, eBioscience), CD45.1 (A20, eBioscience), CD45.2 (104, eBioscience), CD64 (X54-5/7.1, Biolegend), EpCAM (G8.8, eBioscience), CD206 (C068C2, Biolegend), F4/80 (Cl:A3-1, Biorad), Ly6C (HK1.4, eBioscience), Langerin (eBioL31, eBioscience), Ly6G (1A8, BD), MHC II (AF6-120.1, eBioscience), CD9 (MZ3, Biolegend), CD14 (sa2-8, eBioscience), NK1.1 (PK136, eBioscience) and Siglec-F (ES22-10D8, Miltenyi).
Kolter J., Feuerstein R., Zeis P., Hagemeyer N., Paterson N., d'Errico P., Baasch S., Amann L., Masuda T., Lösslein A., Gharun K., Meyer-Luehmann M., Waskow C., Franzke C.W., Grün D., Lämmermann T., Prinz M, & Henneke P. (2019). A Subset of Skin Macrophages Contributes to the Surveillance and Regeneration of Local Nerves. Immunity, 50(6).
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