Cd3e 145 2c11

Manufactured by Thermo Fisher Scientific

Sourced in Austria

The CD3e (145-2C11) is a laboratory reagent used in flow cytometry applications. It is an antibody that binds to the CD3 epsilon chain, a component of the T cell receptor complex. This product is intended for research use only and its core function is to facilitate the identification and analysis of T cells.

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Lab products found in correlation

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Close spelling variants of this product

CD3e (clone 145-2C11; (8 citations) APC-anti-mouse CD3e (145-2C11, (4 citations) Anti-CD3e (145-2C11, (4 citations) PE-labeled anti-CD3e (clone 145-2C11) (2 citations) CD3e (145-2C11)-PE (2 citations) Anti-CD3e (clone 145-2C11, (2 citations) CD3e‐Percp‐cy5.5 (clone 145‐2C11, (2 citations)

8 protocols using cd3e 145 2c11

Flow Cytometric Analysis of Lymphocytes

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Single cell suspensions were prepared from spleen and PLN and stained with fluorescently-labeled antibodies against CD3e (145-2C11), CD4 (GK1.5), CD8a (53–6.7), CD25 (PC61), CD26 (H194-112), CD44 (IM7), CD62L (MEL-14), CD69 (H1.2F3) and matching isotype controls (all eBioscience, San Diego, CA). FoxP3 staining was performed using FoxP3 staining kit (eBioscience). Cells were acquired on a Gallios flow cytometer and data were analyzed with Kaluza software (Beckman Coulter, Suarlée, Belgium).

Ding L., Gysemans C.A., Stangé G., Heremans Y., Yuchi Y., Takiishi T., Korf H., Chintinne M., Carr R.D., Heimberg H., Pipeleers D, & Mathieu C. (2014). Combining MK626, a Novel DPP-4 Inhibitor, and Low-Dose Monoclonal CD3 Antibody for Stable Remission of New-Onset Diabetes in Mice. PLoS ONE, 9(9), e107935.

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Multiparameter Flow Cytometry Analysis

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Antibodies against CD3e (145-2C11), CD4 (GK1.5), NK1.1 (PK136), CD19 (eBio1D3), CD24 (30-F1), CD44 (IM7), CD45.1 (A20), CD45.2 (104), CD48 (HM48-1), CXCR5 (SPRCL5), GL7 (GL-7), GATA3 (TWAJ), IgD (11-26C), ICOS (7E.17G9), Ly9 (Ly9ab3), PD1 (J43), PLZF (Mags.21F7), RORγt (AFKJS-9), 2B4 (eBio244F4), 7-AAD, and streptavidin were purchased from eBioscience. Antibodies against CD138 (281–2) and Fas (Jo2) were purchased from BD. Antibodies against SLAM (12F12), CD84 (CD84.7), and Ly-108 (330-AJ) were purchased from BioLegend, whereas CRACC antibody (clone 520914) was provided by R&D Systems. Antibodies against Egr2 were purchased from Abcam, and goat anti–rabbit IgG (H+L) was purchased from Thermo Fisher Scientific. CD1d-tetramer (19156, loaded with PBS57) and empty CD1d control were kindly provided by the National Institutes of Health Tetramer Core Facility.

Chen S., Cai C., Li Z., Liu G., Wang Y., Blonska M., Li D., Du J., Lin X., Yang M, & Dong Z. (2017). Dissection of SAP-dependent and SAP-independent SLAM family signaling in NKT cell development and humoral immunity. The Journal of Experimental Medicine, 214(2), 475-489.

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Multiparametric Flow Cytometry Analysis

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Cell suspensions from mouse liver, BM (one femur and tibia), spleen, thymus, mesenteric lymph nodes, and peritoneal cavity were processed as previously described (47 (link)), and data were acquired on a FACSCanto10c (BD) and analyzed with FlowJo Software (Tree Star).
The following anti-mouse antibodies were used: B220 (RA3-6B2, BioLegend), BAFFR (7H22-E16,BD), BP-1 (6C3, BioLegend), CD3e (145-2C11, eBioscience), CD4 (GK1.5, eBioscience), CD5 (53-7.3, BioLegend), CD8 (53-6.7, BioLegend), CD9 (MZ3, BioLegend), CD11b (M1/70, eBioscience), CD19 (6D5, 1D3, BioLegend), CD21/CD35 (7E9, BioLegend), CD23 (B3B4, BioLegend), CD24 (M1/69, BioLegend), CD25 (PC61, Biolgend), CD43 (S7, BioLegend), CD44 (IM7, BioLegend), CD45.1 (A20, BioLegend), CD45.2 (104, BioLegend), CD69 (H1.2F3, BioLegend), CD80 (16-10A1, BioLegend), CD86 (GL1, BioLegend), CD93 (AA4.1, BioLegend), DAPI (BIOTIUM), Gr-1 (RB6-8C5, eBioscience), IgDa (AMS-9.1, BioLegend), IgD (11-26c, BioLegend), IgMa (DS-1, MA-69, BioLegend), IgM (Il/41, RMM-1, BioLegend), TCRγδ (GL3, BioLegend), Live dead dye (Zombie Aqua Dye, BioLegend), Live dead dye (Zombie NIR, BioLegend), pERK (4B11B69, BD), pPLCγ2 (K86-1161, BD), Lin28b (AP1485C, ABGENT), and anti-rabbit IgG (BioLegend).

Xu X., Deobagkar-Lele M., Bull K.R., Crockford T.L., Mead A.J., Cribbs A.P., Sims D., Anzilotti C, & Cornall R.J. (2020). An ontogenetic switch drives the positive and negative selection of B cells. Proceedings of the National Academy of Sciences of the United States of America, 117(7), 3718-3727.

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Synthesis and Characterization of Nanoparticles for Immune Modulation

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Nanoparticles were synthesized using DI water, Potassium hexacyanoferrate (II) trihydrate (MW 422.39; K₄[Fe(CN)₆]·3H₂O), iron (III) chloride hexahydrate (MW 270.3; Fe(Cl)₃·6H₂O), Poly (ethylenimine) (PEI, MW 2,000, Mn 1,800, 50% w/v in H₂O), acetate buffer (pH 5.2), citric acid, acetone, and ethanol obtained from Sigma-Aldrich (St. Louis, MO, USA). Murine CpG oligodeoxynucleotide (CpG) TLR9 ligand (ODN 1585; Class A) was purchased from InVivoGen (San Diego, CA, USA). Fluorescent antibodies against HMGB1 (EPR3507) and calreticulin (FMC75) were purchased from Abcam (Cambridge, UK). Dulbecco’s Modified Eagle Media, non-essential amino acids, antibiotic/antimitotic, and β-Mercaptoethanol were purchased from Thermo Fisher (Waltham, Massachusetts). Fetal bovine serum (FBS) was purchased from Atlanta Biologicals (Flowery Branch, GA). CD28 (37.51) and CD3e (145–2C11) antibodies were purchased from eBioscience (San Diego, CA). TexMACS Medium and mouse Pan T Cell Isolation Kit II were purchased from Miltenyi Biotec (Bergisch Gladbach, Germany).

Shukla A., Cano-Mejia J., Andricovich J., Burga R.A., Sweeney E.E, & Fernandes R. (2021). An Engineered Prussian Blue Nanoparticles-based Nanoimmunotherapy Elicits Robust and Persistent Immunological Memory in a TH-MYCN Neuroblastoma Model. Advanced nanobiomed research, 1(8), 2100021.

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Multiparameter Flow Cytometry Analysis

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The following fluorochrome-conjugated anti-mouse antibodies (clones) were used: anti-CD11b (M1/70), -CD11c (N418), -CD45.1 (A20), -CD80 (16-10A1), -CD86 (GL-1), and -CD4 (RM4-5) all from BioLegend (San Diego, CA); anti-GR1 (RB6-8C5), -Ly6C (HK1.4), -CD3e (145-2C11), and -CD28 (37.51) all from eBioscience (San Diego, CA); and anti-MHC-II (M5/114.15.2) from BD Bioscience (San Jose, CA). LIVE/DEAD stain was used to determine the viability of cells and was purchased from Life Technologies (Eugene, OR). Flow cytometry was performed using an LSRII instrument and the acquired data were analyzed using FlowJo software.

Avery J.T., Jimenez R.V., Blake J.L., Wright T.T., Leόn-Ruiz B., Schoeb T.R., Szalai A.J, & Bullard D.C. (2019). Mice expressing the variant rs1143679 allele of ITGAM (CD11b) show impaired DC-mediated T cell proliferation. Mammalian Genome, 30(9), 245-259.

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Multicolor Flow Cytometry Analysis of Murine Splenocytes

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Spleen single-cell suspensions were incubated with a Fc-receptor blocking antibody 2.4G2 (kindly provided by Dr. Louis Boon, Bioceros BV, Utrecht, The Netherlands) and the percentage of spleen cell populations (macrophages, B cells, Dendritic cells, T cells, monocytes) were determined using the following antibodies: F4/80 (BM8) (Invitrogen, Bleiswijk, The Netherlands), CD45R (B220) (eBioscience, Vienna, Austria), CD11c (HL3) (BD Biosciences), CD4 (GK1.5) (eBioscience, Vienna, Austria), CD8α (53-6.7) (eBioscience, Vienna, Austria), CD3e (145-2C11) (eBioscience, Vienna, Austria), Lineage (Lin) (B220, NK1.1, CD90, CD49. Ly6G) (ebioscience, Vienna, Austria) CD45.1 (ebioscience, Vienna, Austria). Samples were analyzed with a LSR Fortessa II (Beckman Coulter) and the FlowJo software (Tree Star Inc., Ashland, The United States). The different spleen cell populations were defined as follow: DCs (CD11c+MHCII+), macrophages (F4/80+), B cells (CD45R+MHCII+), CD4+ (CD3+CD8-CD4+), CD8+T cells (CD3+CD8+CD4-) and monocytes (CD45+Lin-F4/80-CD11c-MHCII-CD11b+Ly-6C+). Percentages were reported to the total number of splenocytes of each mouse to calculate the number of cells per population (Figure S2 and S3).

Costes L.M., van der Vliet J., Farro G., Matteoli G., van Bree S.H., Olivier B.J., Nolte M.A., Boeckxstaens G.E, & Cailotto C. (2014). The Spleen Responds to Intestinal Manipulation but Does Not Participate in the Inflammatory Response in a Mouse Model of Postoperative Ileus. PLoS ONE, 9(7), e102211.

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Flow Cytometry Immunophenotyping Protocol

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The following antibodies were used for flow cytometry: CD3e (145-2C11; eBioscience and 500A2; BD), CD4 (RM4-5; BD), CD45.2 (104; eBioscience), CD90.1 (HIS51; eBioscience). Intracellular cytokine staining was performed with anti-IFNγ (XMG1.2; BD) or anti-TNFα (MP6-XT22; BD), using the Cytofix/Cytoperm kit (BD Biosciences). Flow cytometric data were collected on an LSR II (BD Biosciences) and analyzed with FlowJo software (TreeStar).

Torabi-Parizi P., Vrisekoop N., Kastenmuller W., Gerner M.Y., Egen J.G, & Germain R.N. (2014). Pathogen-related differences in the abundance of presented antigen are reflected in CD4+ T cell dynamic behavior and effector function in the lung. Journal of immunology (Baltimore, Md. : 1950), 192(4), 1651-1660.

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Isolation and Characterization of Murine Hematopoietic Stem and Progenitor Cells

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Bone marrow cells were isolated by flushing one femur and tibia with 3 mL Ca2+ and Mg2+ free HBSS supplemented with 2% HI FBS, filtered through a 50 μm mesh and counted using a hemocytometer. Splenocytes were obtained by mashing between two glass slides and filtered through a 100 μm mesh. Lineage cocktail was comprised of biotin labeled lineage panel (CD3e 145-2C11, B220 RA3-6B2, TER119, Gr-1 RB6-8C5, Mac1 M1/70 eBioscience) along with biotin CD4 (GK1.5 BioLegend) and CD8a (53-6.7 BioLegend) followed by staining with Streptavidin Pacific Orange (Invitrogen). Antibodies used to stain HSPCs included PE CD150 (TC15-12F123.2 BioLegend), PE Cy7 CD48 (HM48-1 BioLegend), PE Cy7 CD41 (MWReg eBiosciences), APC Sca1 (E13-161.7 BioLegend), APC Cy7 cKit (2B8 BioLegend). Antibodies used to stain peripheral blood included: FITC CD45.2 (104 BioLegend), PE CD3e (145-2C11 BioLegend), PE CD115 (AFS98 BioLegend), PE Cy5 B220 (RA3-6B2 eBioscience), PE Cy7 Gr-1 (RB6-8C5 BioLegend), PE Cy7 Ly6G (1A8 BD Pharmingen), APC Mac1 (M1/70 eBioscience), APC Cy7 CD45.1 (A20 eBioscience), biotin CD3e (145-2C11, eBioscience), biotin CD4 (GK1.5 BioLegend) and biotin CD8a (53-6.7 BioLegend).

Burberry A., Zeng M.Y., Ding L., Wicks I., Inohara N., Morrison S.J, & Núñez G. (2014). Infection Mobilizes Hematopoietic Stem Cells through Cooperative NOD-like Receptor and Toll-like Receptor Signaling. Cell host & microbe, 15(6), 779-791.

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