cd3e functional grade monoclonal antibody  (Thermo Fisher)


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    CD3e Functional Grade Monoclonal Antibody 145 2C11
    Description:
    CD3e Functional Grade Monoclonal Antibody for Flow FN
    Catalog Number:
    16-0031-81
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    Category:
    Antibodies Secondary Detection Reagents
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    Structured Review

    Thermo Fisher cd3e functional grade monoclonal antibody
    Impact of Nf1 heterozygosity on T cells proliferation and function. a , b Proliferation of peripheral T cells from Nf1 +/+ and Nf1 +/− mice stimulated by various concentrations of a <t>anti-CD3e</t> antibody alone or with b CD28 costimulation. Proliferation was assessed in biological duplicate and technical triplicate by 3 H-thymidine incorporation during the last 20 h of culture. c Delayed-type hypersensitivity (DTH) assay: On day 0, Nf1 +/+ and Nf1 +/− mice were sensitized on abdominal skin by topical application of oxazolone. On day 6, Nf1 +/+ and Nf1 +/− mice were challenged by topical application on the left and right ear with oxazolone and control solvent, respectively. DTH was assessed daily through by measuring ear thickness (Δ). Experiments were performed in biological triplicates. d Two days after challenge, cellularity of draining lymph nodes (DLNs) of Nf1 +/+ and Nf1 +/− mice were determined. e Spontaneous activation of cells from DLNs (T cells) was measured by 3 H-thymidine incorporation after culturing for 3 days without stimuli. f Fraction of CD4 + cells that are activated. g Fraction of CD8 + cells that are activated. h Fraction of B cells + that are activated. i Fraction of Treg + cells in total CD4 + subset. Panels d – i were performed in biological triplicates. Statistical paired t -test performed; * P
    CD3e Functional Grade Monoclonal Antibody for Flow FN
    https://www.bioz.com/result/cd3e functional grade monoclonal antibody/product/Thermo Fisher
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cd3e functional grade monoclonal antibody - by Bioz Stars, 2021-05
    94/100 stars

    Images

    1) Product Images from "NF1 heterozygosity fosters de novo tumorigenesis but impairs malignant transformation"

    Article Title: NF1 heterozygosity fosters de novo tumorigenesis but impairs malignant transformation

    Journal: Nature Communications

    doi: 10.1038/s41467-018-07452-y

    Impact of Nf1 heterozygosity on T cells proliferation and function. a , b Proliferation of peripheral T cells from Nf1 +/+ and Nf1 +/− mice stimulated by various concentrations of a anti-CD3e antibody alone or with b CD28 costimulation. Proliferation was assessed in biological duplicate and technical triplicate by 3 H-thymidine incorporation during the last 20 h of culture. c Delayed-type hypersensitivity (DTH) assay: On day 0, Nf1 +/+ and Nf1 +/− mice were sensitized on abdominal skin by topical application of oxazolone. On day 6, Nf1 +/+ and Nf1 +/− mice were challenged by topical application on the left and right ear with oxazolone and control solvent, respectively. DTH was assessed daily through by measuring ear thickness (Δ). Experiments were performed in biological triplicates. d Two days after challenge, cellularity of draining lymph nodes (DLNs) of Nf1 +/+ and Nf1 +/− mice were determined. e Spontaneous activation of cells from DLNs (T cells) was measured by 3 H-thymidine incorporation after culturing for 3 days without stimuli. f Fraction of CD4 + cells that are activated. g Fraction of CD8 + cells that are activated. h Fraction of B cells + that are activated. i Fraction of Treg + cells in total CD4 + subset. Panels d – i were performed in biological triplicates. Statistical paired t -test performed; * P
    Figure Legend Snippet: Impact of Nf1 heterozygosity on T cells proliferation and function. a , b Proliferation of peripheral T cells from Nf1 +/+ and Nf1 +/− mice stimulated by various concentrations of a anti-CD3e antibody alone or with b CD28 costimulation. Proliferation was assessed in biological duplicate and technical triplicate by 3 H-thymidine incorporation during the last 20 h of culture. c Delayed-type hypersensitivity (DTH) assay: On day 0, Nf1 +/+ and Nf1 +/− mice were sensitized on abdominal skin by topical application of oxazolone. On day 6, Nf1 +/+ and Nf1 +/− mice were challenged by topical application on the left and right ear with oxazolone and control solvent, respectively. DTH was assessed daily through by measuring ear thickness (Δ). Experiments were performed in biological triplicates. d Two days after challenge, cellularity of draining lymph nodes (DLNs) of Nf1 +/+ and Nf1 +/− mice were determined. e Spontaneous activation of cells from DLNs (T cells) was measured by 3 H-thymidine incorporation after culturing for 3 days without stimuli. f Fraction of CD4 + cells that are activated. g Fraction of CD8 + cells that are activated. h Fraction of B cells + that are activated. i Fraction of Treg + cells in total CD4 + subset. Panels d – i were performed in biological triplicates. Statistical paired t -test performed; * P

    Techniques Used: Mouse Assay, Activation Assay

    2) Product Images from "NF1 heterozygosity fosters de novo tumorigenesis but impairs malignant transformation"

    Article Title: NF1 heterozygosity fosters de novo tumorigenesis but impairs malignant transformation

    Journal: Nature Communications

    doi: 10.1038/s41467-018-07452-y

    Impact of Nf1 heterozygosity on T cells proliferation and function. a , b Proliferation of peripheral T cells from Nf1 +/+ and Nf1 +/− mice stimulated by various concentrations of a anti-CD3e antibody alone or with b CD28 costimulation. Proliferation was assessed in biological duplicate and technical triplicate by 3 H-thymidine incorporation during the last 20 h of culture. c Delayed-type hypersensitivity (DTH) assay: On day 0, Nf1 +/+ and Nf1 +/− mice were sensitized on abdominal skin by topical application of oxazolone. On day 6, Nf1 +/+ and Nf1 +/− mice were challenged by topical application on the left and right ear with oxazolone and control solvent, respectively. DTH was assessed daily through by measuring ear thickness (Δ). Experiments were performed in biological triplicates. d Two days after challenge, cellularity of draining lymph nodes (DLNs) of Nf1 +/+ and Nf1 +/− mice were determined. e Spontaneous activation of cells from DLNs (T cells) was measured by 3 H-thymidine incorporation after culturing for 3 days without stimuli. f Fraction of CD4 + cells that are activated. g Fraction of CD8 + cells that are activated. h Fraction of B cells + that are activated. i Fraction of Treg + cells in total CD4 + subset. Panels d – i were performed in biological triplicates. Statistical paired t -test performed; * P
    Figure Legend Snippet: Impact of Nf1 heterozygosity on T cells proliferation and function. a , b Proliferation of peripheral T cells from Nf1 +/+ and Nf1 +/− mice stimulated by various concentrations of a anti-CD3e antibody alone or with b CD28 costimulation. Proliferation was assessed in biological duplicate and technical triplicate by 3 H-thymidine incorporation during the last 20 h of culture. c Delayed-type hypersensitivity (DTH) assay: On day 0, Nf1 +/+ and Nf1 +/− mice were sensitized on abdominal skin by topical application of oxazolone. On day 6, Nf1 +/+ and Nf1 +/− mice were challenged by topical application on the left and right ear with oxazolone and control solvent, respectively. DTH was assessed daily through by measuring ear thickness (Δ). Experiments were performed in biological triplicates. d Two days after challenge, cellularity of draining lymph nodes (DLNs) of Nf1 +/+ and Nf1 +/− mice were determined. e Spontaneous activation of cells from DLNs (T cells) was measured by 3 H-thymidine incorporation after culturing for 3 days without stimuli. f Fraction of CD4 + cells that are activated. g Fraction of CD8 + cells that are activated. h Fraction of B cells + that are activated. i Fraction of Treg + cells in total CD4 + subset. Panels d – i were performed in biological triplicates. Statistical paired t -test performed; * P

    Techniques Used: Mouse Assay, Activation Assay

    Related Articles

    Purification:

    Article Title: Epigallocatechin-3-gallate (EGCG) up-regulates miR-15b expression thus attenuating store operated calcium entry (SOCE) into murine CD4+ T cells and human leukaemic T cell lymphoblasts
    Article Snippet: Murine CD4+ T cells isolation and culture and treatment Murine CD4+ naive T cells were isolated from C57BL/6 mice using the MagniSort® Mouse naïve T cell Enrichment kit (#8804-6824-74, eBioscience, Germany) as described by the manufacturer. .. Purified T cells were cultured in plate-bound anti-CD3 (#16-0031-85, eBioscience)/anti-CD28 (#16-0281-85, eBioscience) Abs at a 1:2 ratio (1μg/ml anti-CD3 and 2μg/ml anti-CD28) in the presence or absence of 5-50 μM EGCG (#E4143, Sigma, Germany) for 3 days. .. Human leukaemic T cell lymphoblasts (Jurkat T cells) culture and treatment Human Jurkat E6.1 Cell Line (#88042803, Sigma) were maintained in RPMI 1640 (#61870-010, Life Technologies, Germany) medium supplemented with 10% Fetal bovine serum (#10270-106, Life Technologies), 1% penicillin/streptomycin (#P4333, Sigma) and 0.1% 2-Mercaptoethanol (#31350-010, Life Technologies) in cell culture flasks, then incubated at 37°C in a humidified atmosphere at 5% (v/v) CO2 .

    Article Title: NF1 heterozygosity fosters de novo tumorigenesis but impairs malignant transformation
    Article Snippet: T cell proliferation assay Following the manufacturer’s recommendations, CD3+ T cells were purified from the lymph nodes of Nf1 +/+ and Nf1 +/− mice (age 3–6 months) using pan-T-cell isolation kit (Miltenyi Biotec, Auburn, CA). .. Purified CD3+ T cells (2 × 105 /well) were cultured for 2 days in 96-well plates (in triplicate) precoated with indicated doses of anti-CD3 Ab (eBioscience; cat. no. 16-0031-86) alone or anti-CD28 mAb (eBioscience; cat. no. 16-0281-85). .. After pulsing with 3 H-thymidine (1 μCi/well [0.037 MBq/well]) for 20–22 h, cells were collected and evaluated for 3 H radioactivity.

    Cell Culture:

    Article Title: Epigallocatechin-3-gallate (EGCG) up-regulates miR-15b expression thus attenuating store operated calcium entry (SOCE) into murine CD4+ T cells and human leukaemic T cell lymphoblasts
    Article Snippet: Murine CD4+ T cells isolation and culture and treatment Murine CD4+ naive T cells were isolated from C57BL/6 mice using the MagniSort® Mouse naïve T cell Enrichment kit (#8804-6824-74, eBioscience, Germany) as described by the manufacturer. .. Purified T cells were cultured in plate-bound anti-CD3 (#16-0031-85, eBioscience)/anti-CD28 (#16-0281-85, eBioscience) Abs at a 1:2 ratio (1μg/ml anti-CD3 and 2μg/ml anti-CD28) in the presence or absence of 5-50 μM EGCG (#E4143, Sigma, Germany) for 3 days. .. Human leukaemic T cell lymphoblasts (Jurkat T cells) culture and treatment Human Jurkat E6.1 Cell Line (#88042803, Sigma) were maintained in RPMI 1640 (#61870-010, Life Technologies, Germany) medium supplemented with 10% Fetal bovine serum (#10270-106, Life Technologies), 1% penicillin/streptomycin (#P4333, Sigma) and 0.1% 2-Mercaptoethanol (#31350-010, Life Technologies) in cell culture flasks, then incubated at 37°C in a humidified atmosphere at 5% (v/v) CO2 .

    Article Title: NF1 heterozygosity fosters de novo tumorigenesis but impairs malignant transformation
    Article Snippet: T cell proliferation assay Following the manufacturer’s recommendations, CD3+ T cells were purified from the lymph nodes of Nf1 +/+ and Nf1 +/− mice (age 3–6 months) using pan-T-cell isolation kit (Miltenyi Biotec, Auburn, CA). .. Purified CD3+ T cells (2 × 105 /well) were cultured for 2 days in 96-well plates (in triplicate) precoated with indicated doses of anti-CD3 Ab (eBioscience; cat. no. 16-0031-86) alone or anti-CD28 mAb (eBioscience; cat. no. 16-0281-85). .. After pulsing with 3 H-thymidine (1 μCi/well [0.037 MBq/well]) for 20–22 h, cells were collected and evaluated for 3 H radioactivity.

    Article Title: CD1b-autoreactive T cells recognize phospholipid antigens and contribute to antitumor immunity against a CD1b+ T cell lymphoma
    Article Snippet: .. Lymphocytes isolated from the spleen and lymph nodes of HJ1Tg/hCD1Tg/Rag2−/− mice were stimulated with 5 μg/mL anti-CD3 (eBioscience, 16-0031-86), 2 μg/mL anti-CD28 (Bioxcell, BE00155), and 20 ng/mL IL-12 (Peprotech, 210–12) for 48 h. The cells were “rested” for 48 h in 70% RPMI-10 medium and 30% supplemented Mischell Dutton media with 20 U/mL IL-2 (Peprotech, 212–12) (CTL medium) and cultured for an additional 3 d in 100% CTL medium. ..

    Activation Assay:

    Article Title: MDM2 inhibitor APG-115 synergizes with PD-1 blockade through enhancing antitumor immunity in the tumor microenvironment
    Article Snippet: .. Analyses of T cell activation and proliferation CD4+ T cells were positively selected from mouse spleens using magnetic beads (Miltenyi, catalog #130–049-201) and stimulated with 10 μg/mL plate-bound anti-CD3 (eBioscience, catalog #16–0031-85) and 2 μg/mL anti-CD28 (eBioscience, catalog #16–0281-85) in the presence of 250 nM APG-115 or DMSO for 1 or 2 days. .. After treatment, cells were harvested and evaluated by flow cytometry for the expression of CD25 (BD, catalog #557192), CD62L (BD, catalog #553151), and Foxp3 (Thermo Fisher, catalog #12–5773-82).

    Magnetic Beads:

    Article Title: MDM2 inhibitor APG-115 synergizes with PD-1 blockade through enhancing antitumor immunity in the tumor microenvironment
    Article Snippet: .. Analyses of T cell activation and proliferation CD4+ T cells were positively selected from mouse spleens using magnetic beads (Miltenyi, catalog #130–049-201) and stimulated with 10 μg/mL plate-bound anti-CD3 (eBioscience, catalog #16–0031-85) and 2 μg/mL anti-CD28 (eBioscience, catalog #16–0281-85) in the presence of 250 nM APG-115 or DMSO for 1 or 2 days. .. After treatment, cells were harvested and evaluated by flow cytometry for the expression of CD25 (BD, catalog #557192), CD62L (BD, catalog #553151), and Foxp3 (Thermo Fisher, catalog #12–5773-82).

    Flow Cytometry:

    Article Title: MFG-E8 Is Critical for Embryonic Stem Cell-Mediated T Cell Immunomodulation
    Article Snippet: After two washes, splenocytes were re-suspended, and CD3+ T cells ( > 92% purity) or NK cells ( > 90% purity) were fractionated by negative selection using a magnetic labeling kit according to manufacturer instructions (STEMCELL Technologies, catalog no. 19851). .. Flow Cytometry and Intracellular Staining Fractionated CD3+ T cells were treated with different reagents as described in , followed by stimulation with plate-coated anti-CD3 and anti-CD28 (eBioscience, catalog no. 16-0031-85 and 16-0281-85) for 48 hr. .. Cells were then stained with antibodies against CD4, CD8, and surface activation markers (CD25, CD44, and CD69) (eBioscience, catalog no. 15-0041-82, 11-0081-81, 48-0253-82, 25-0691-81, and 47-0441-82, respectively).

    Cytometry:

    Article Title: MFG-E8 Is Critical for Embryonic Stem Cell-Mediated T Cell Immunomodulation
    Article Snippet: After two washes, splenocytes were re-suspended, and CD3+ T cells ( > 92% purity) or NK cells ( > 90% purity) were fractionated by negative selection using a magnetic labeling kit according to manufacturer instructions (STEMCELL Technologies, catalog no. 19851). .. Flow Cytometry and Intracellular Staining Fractionated CD3+ T cells were treated with different reagents as described in , followed by stimulation with plate-coated anti-CD3 and anti-CD28 (eBioscience, catalog no. 16-0031-85 and 16-0281-85) for 48 hr. .. Cells were then stained with antibodies against CD4, CD8, and surface activation markers (CD25, CD44, and CD69) (eBioscience, catalog no. 15-0041-82, 11-0081-81, 48-0253-82, 25-0691-81, and 47-0441-82, respectively).

    Staining:

    Article Title: MFG-E8 Is Critical for Embryonic Stem Cell-Mediated T Cell Immunomodulation
    Article Snippet: After two washes, splenocytes were re-suspended, and CD3+ T cells ( > 92% purity) or NK cells ( > 90% purity) were fractionated by negative selection using a magnetic labeling kit according to manufacturer instructions (STEMCELL Technologies, catalog no. 19851). .. Flow Cytometry and Intracellular Staining Fractionated CD3+ T cells were treated with different reagents as described in , followed by stimulation with plate-coated anti-CD3 and anti-CD28 (eBioscience, catalog no. 16-0031-85 and 16-0281-85) for 48 hr. .. Cells were then stained with antibodies against CD4, CD8, and surface activation markers (CD25, CD44, and CD69) (eBioscience, catalog no. 15-0041-82, 11-0081-81, 48-0253-82, 25-0691-81, and 47-0441-82, respectively).

    Recombinant:

    Article Title: Pulmonary T cell activation in response to chronic particulate air pollution
    Article Snippet: Cells were incubated at 37°C for 10 min followed by two washes of 1 ml of RPMI with 10% FBS and then spun in a microcentrifuge at 250 g for 5 min. T cells were resuspended in T cell growth media before seeding at 2 × 105 cells/well of a 96-well plate with confluent, pretreated BMM (see above). .. T cell growth media containing 2 μg/ml anti-CD3 (16–0031-85 clone 145–2c11; eBiosciences) antibody was added to RPMI with 20% FBS. recombinant mouse macrophage colony-stimulating factor (R & D systems), 10 ng/ml, was added to the T cell growth media whenever T cells were cocultured with BMMs. .. After 72 h of culture, T cells were run on a flow cytometer, and proliferation index was calculated according to the equation outlined by Wallace and Muirhead ( ).

    Isolation:

    Article Title: CD1b-autoreactive T cells recognize phospholipid antigens and contribute to antitumor immunity against a CD1b+ T cell lymphoma
    Article Snippet: .. Lymphocytes isolated from the spleen and lymph nodes of HJ1Tg/hCD1Tg/Rag2−/− mice were stimulated with 5 μg/mL anti-CD3 (eBioscience, 16-0031-86), 2 μg/mL anti-CD28 (Bioxcell, BE00155), and 20 ng/mL IL-12 (Peprotech, 210–12) for 48 h. The cells were “rested” for 48 h in 70% RPMI-10 medium and 30% supplemented Mischell Dutton media with 20 U/mL IL-2 (Peprotech, 212–12) (CTL medium) and cultured for an additional 3 d in 100% CTL medium. ..

    Mouse Assay:

    Article Title: CD1b-autoreactive T cells recognize phospholipid antigens and contribute to antitumor immunity against a CD1b+ T cell lymphoma
    Article Snippet: .. Lymphocytes isolated from the spleen and lymph nodes of HJ1Tg/hCD1Tg/Rag2−/− mice were stimulated with 5 μg/mL anti-CD3 (eBioscience, 16-0031-86), 2 μg/mL anti-CD28 (Bioxcell, BE00155), and 20 ng/mL IL-12 (Peprotech, 210–12) for 48 h. The cells were “rested” for 48 h in 70% RPMI-10 medium and 30% supplemented Mischell Dutton media with 20 U/mL IL-2 (Peprotech, 212–12) (CTL medium) and cultured for an additional 3 d in 100% CTL medium. ..

    CTL Assay:

    Article Title: CD1b-autoreactive T cells recognize phospholipid antigens and contribute to antitumor immunity against a CD1b+ T cell lymphoma
    Article Snippet: .. Lymphocytes isolated from the spleen and lymph nodes of HJ1Tg/hCD1Tg/Rag2−/− mice were stimulated with 5 μg/mL anti-CD3 (eBioscience, 16-0031-86), 2 μg/mL anti-CD28 (Bioxcell, BE00155), and 20 ng/mL IL-12 (Peprotech, 210–12) for 48 h. The cells were “rested” for 48 h in 70% RPMI-10 medium and 30% supplemented Mischell Dutton media with 20 U/mL IL-2 (Peprotech, 212–12) (CTL medium) and cultured for an additional 3 d in 100% CTL medium. ..

    Positive Control:

    Article Title: Blocking Stroke-Induced Immunodeficiency Increases CNS Antigen-Specific Autoreactivity But Does Not Worsen Functional Outcome after Experimental Stroke
    Article Snippet: Plates were washed with sterile PBS and blocked with 1% BSA fraction V (Sigma-Aldrich) in PBS for 2 h at room temperature. .. Wells were filled with 100 μl of the following: (1) complete HL-1 medium (Lonza) supplemented with 2 m m l -alanyl- l -glutamine, 50 U/ml penicillin, and 50 μg/ml streptomycin, with 2 μg/ml CD-3 (145-2C11; eBioscience, catalog #16-0031-86; RRID:AB_468849) and 0.2 μg/ml CD-28 (37.51; eBioscience, catalog #14-0281-85; RRID:AB_467191); duplicates per sample (positive control); (2) complete HL-1 medium with 0.2 μg/ml CD-28 and 40 μg/ml myelin oligodendrocyte glycoprotein peptide (pMOG35–55 , amino acid sequence: MEVGWYRSPFSRVVHLYRNGK; a gift from Dr. Rudolf Volkmer; Charité Universitätsmedizin Berlin, Berlin, Germany); triplicates per sample (MOG35–55 -peptide stimulation); and (3) complete HL-1 medium with 0.2 μg/ml CD-28; triplicates per sample (negative control). .. In brain Elispot, brain MNCs were added at 1.75 × 105 in 50 μl complete HL-1 medium per well together with 1 × 106 naive C57BL/6J splenocytes in 50 μl complete HL-1 medium.

    Sequencing:

    Article Title: Blocking Stroke-Induced Immunodeficiency Increases CNS Antigen-Specific Autoreactivity But Does Not Worsen Functional Outcome after Experimental Stroke
    Article Snippet: Plates were washed with sterile PBS and blocked with 1% BSA fraction V (Sigma-Aldrich) in PBS for 2 h at room temperature. .. Wells were filled with 100 μl of the following: (1) complete HL-1 medium (Lonza) supplemented with 2 m m l -alanyl- l -glutamine, 50 U/ml penicillin, and 50 μg/ml streptomycin, with 2 μg/ml CD-3 (145-2C11; eBioscience, catalog #16-0031-86; RRID:AB_468849) and 0.2 μg/ml CD-28 (37.51; eBioscience, catalog #14-0281-85; RRID:AB_467191); duplicates per sample (positive control); (2) complete HL-1 medium with 0.2 μg/ml CD-28 and 40 μg/ml myelin oligodendrocyte glycoprotein peptide (pMOG35–55 , amino acid sequence: MEVGWYRSPFSRVVHLYRNGK; a gift from Dr. Rudolf Volkmer; Charité Universitätsmedizin Berlin, Berlin, Germany); triplicates per sample (MOG35–55 -peptide stimulation); and (3) complete HL-1 medium with 0.2 μg/ml CD-28; triplicates per sample (negative control). .. In brain Elispot, brain MNCs were added at 1.75 × 105 in 50 μl complete HL-1 medium per well together with 1 × 106 naive C57BL/6J splenocytes in 50 μl complete HL-1 medium.

    Negative Control:

    Article Title: Blocking Stroke-Induced Immunodeficiency Increases CNS Antigen-Specific Autoreactivity But Does Not Worsen Functional Outcome after Experimental Stroke
    Article Snippet: Plates were washed with sterile PBS and blocked with 1% BSA fraction V (Sigma-Aldrich) in PBS for 2 h at room temperature. .. Wells were filled with 100 μl of the following: (1) complete HL-1 medium (Lonza) supplemented with 2 m m l -alanyl- l -glutamine, 50 U/ml penicillin, and 50 μg/ml streptomycin, with 2 μg/ml CD-3 (145-2C11; eBioscience, catalog #16-0031-86; RRID:AB_468849) and 0.2 μg/ml CD-28 (37.51; eBioscience, catalog #14-0281-85; RRID:AB_467191); duplicates per sample (positive control); (2) complete HL-1 medium with 0.2 μg/ml CD-28 and 40 μg/ml myelin oligodendrocyte glycoprotein peptide (pMOG35–55 , amino acid sequence: MEVGWYRSPFSRVVHLYRNGK; a gift from Dr. Rudolf Volkmer; Charité Universitätsmedizin Berlin, Berlin, Germany); triplicates per sample (MOG35–55 -peptide stimulation); and (3) complete HL-1 medium with 0.2 μg/ml CD-28; triplicates per sample (negative control). .. In brain Elispot, brain MNCs were added at 1.75 × 105 in 50 μl complete HL-1 medium per well together with 1 × 106 naive C57BL/6J splenocytes in 50 μl complete HL-1 medium.

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    Thermo Fisher anti cd3
    T regulatory cells (Tregs) increase in no. and activation state centrally but not in the lung. GFP, green fluorescent protein. A : total <t>CD3</t> + , CD3 + CD4 + , and CD3 + CD8 + cells in the lung of GFP. Foxp3.KI mice responded to PM 2.5 exposure similar to the WT
    Anti Cd3, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cd3/product/Thermo Fisher
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cd3 - by Bioz Stars, 2021-05
    94/100 stars
      Buy from Supplier

    Image Search Results


    T regulatory cells (Tregs) increase in no. and activation state centrally but not in the lung. GFP, green fluorescent protein. A : total CD3 + , CD3 + CD4 + , and CD3 + CD8 + cells in the lung of GFP. Foxp3.KI mice responded to PM 2.5 exposure similar to the WT

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    Article Title: Pulmonary T cell activation in response to chronic particulate air pollution

    doi: 10.1152/ajplung.00261.2011

    Figure Lengend Snippet: T regulatory cells (Tregs) increase in no. and activation state centrally but not in the lung. GFP, green fluorescent protein. A : total CD3 + , CD3 + CD4 + , and CD3 + CD8 + cells in the lung of GFP. Foxp3.KI mice responded to PM 2.5 exposure similar to the WT

    Article Snippet: T cell growth media containing 2 μg/ml anti-CD3 (16–0031-85 clone 145–2c11; eBiosciences) antibody was added to RPMI with 20% FBS. recombinant mouse macrophage colony-stimulating factor (R & D systems), 10 ng/ml, was added to the T cell growth media whenever T cells were cocultured with BMMs.

    Techniques: Activation Assay, Mouse Assay

    PM 2.5 potentiates T cell infiltration, activation, and CXCR3 expression in the lung. A : WT-PM 2.5 mice had significantly more CD3 + cells/g lung than WT-FA. CD3 + CD4 + cells increased significantly in WT-PM 2.5 mice ( P = 0.007) but not in CXCR3 −/−

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    Article Title: Pulmonary T cell activation in response to chronic particulate air pollution

    doi: 10.1152/ajplung.00261.2011

    Figure Lengend Snippet: PM 2.5 potentiates T cell infiltration, activation, and CXCR3 expression in the lung. A : WT-PM 2.5 mice had significantly more CD3 + cells/g lung than WT-FA. CD3 + CD4 + cells increased significantly in WT-PM 2.5 mice ( P = 0.007) but not in CXCR3 −/−

    Article Snippet: T cell growth media containing 2 μg/ml anti-CD3 (16–0031-85 clone 145–2c11; eBiosciences) antibody was added to RPMI with 20% FBS. recombinant mouse macrophage colony-stimulating factor (R & D systems), 10 ng/ml, was added to the T cell growth media whenever T cells were cocultured with BMMs.

    Techniques: Activation Assay, Expressing, Mouse Assay

    Flow cytometry analysis of TILs in the TME of syngeneic tumors with wild-type ( a ) or mutant ( b ) Trp53 . Mice with established MH-22A or MC38 tumors were treated with 10 mg/kg APG-115 ( a and b ), 10 mg/kg ( a ) or 5 mg/kg ( b ) anti-PD-1 antibody, or the combination as described in the legend of Fig. 4 . The control group was treated with isotype control antibody and APG-115 vehicle (I + V). On day 14, syngeneic tumors were harvested, dissociated into single-cell suspensions, and stained for flow cytometry analysis. Percentages of CD45 + , CD3 + T cells, CD8 + T cells, M1 and M2 macrophages in the tumors under the different treatments were assessed. Data were representative of two ( a ) or three ( b ) independent experiments and shown as dot plots ( n = 5 or 10). **** P

    Journal: Journal for Immunotherapy of Cancer

    Article Title: MDM2 inhibitor APG-115 synergizes with PD-1 blockade through enhancing antitumor immunity in the tumor microenvironment

    doi: 10.1186/s40425-019-0750-6

    Figure Lengend Snippet: Flow cytometry analysis of TILs in the TME of syngeneic tumors with wild-type ( a ) or mutant ( b ) Trp53 . Mice with established MH-22A or MC38 tumors were treated with 10 mg/kg APG-115 ( a and b ), 10 mg/kg ( a ) or 5 mg/kg ( b ) anti-PD-1 antibody, or the combination as described in the legend of Fig. 4 . The control group was treated with isotype control antibody and APG-115 vehicle (I + V). On day 14, syngeneic tumors were harvested, dissociated into single-cell suspensions, and stained for flow cytometry analysis. Percentages of CD45 + , CD3 + T cells, CD8 + T cells, M1 and M2 macrophages in the tumors under the different treatments were assessed. Data were representative of two ( a ) or three ( b ) independent experiments and shown as dot plots ( n = 5 or 10). **** P

    Article Snippet: Analyses of T cell activation and proliferation CD4+ T cells were positively selected from mouse spleens using magnetic beads (Miltenyi, catalog #130–049-201) and stimulated with 10 μg/mL plate-bound anti-CD3 (eBioscience, catalog #16–0031-85) and 2 μg/mL anti-CD28 (eBioscience, catalog #16–0281-85) in the presence of 250 nM APG-115 or DMSO for 1 or 2 days.

    Techniques: Flow Cytometry, Cytometry, Mutagenesis, Mouse Assay, Staining

    APG-115 suppresses alternative M2 macrophages polarization in vitro and increases M1 macrophages in vivo through activation of p53 pathway. a BMDMs were generated under the treatment with m-CSF for 7 days and then treated with IL-4 (20 ng/mL) to induce alternative macrophage polarization (M2) for 24 h in the absence or presence of APG-115. Cells were then harvested for detection of M2 macrophages (CD206 + MHC-II low ) by flow cytometry. b the mRNA expression levels of Arg-1 and Retnla in the above BMDMs induced by the treatment with IL-4 (20 ng/mL) with or without APG-115 were analyzed by RT-qPCR. Duplicated samples were tested. c Western blot analysis of p53, p21, c-Myc and c-Maf total proteins in BMDMs treated with IL-4 (20 ng/mL) with or without APG-115 (1 μM) for 0, 4, or 24 h, or sequentially treated with IL-4 and then APG-115 for 24 h each (24 h + 24 h). d Quantification of C. 0 (black bars), 4 (blue bars), or 24 h (green bars), or sequentially treated with each agent for 24 h (24 h + 24 h, red bars). e naïve BALB/c mice were treated with APG-115 (10 mg/kg, Q2D × 2 doses; n = 5). Two days after the last dose, spleens were collected, dissociated into single-cell suspensions, and stained with macrophage markers for flow cytometry analysis. Macrophages were defined as CD11b + F4/80 hi , and further analyzed for M1 macrophages by expression of MHC-II. Pooled data of percentages of macrophages gated on CD45 + CD3 − live cells ( f ) and percentages of M1 macrophages gated on macrophages ( g ) from five mice were plotted

    Journal: Journal for Immunotherapy of Cancer

    Article Title: MDM2 inhibitor APG-115 synergizes with PD-1 blockade through enhancing antitumor immunity in the tumor microenvironment

    doi: 10.1186/s40425-019-0750-6

    Figure Lengend Snippet: APG-115 suppresses alternative M2 macrophages polarization in vitro and increases M1 macrophages in vivo through activation of p53 pathway. a BMDMs were generated under the treatment with m-CSF for 7 days and then treated with IL-4 (20 ng/mL) to induce alternative macrophage polarization (M2) for 24 h in the absence or presence of APG-115. Cells were then harvested for detection of M2 macrophages (CD206 + MHC-II low ) by flow cytometry. b the mRNA expression levels of Arg-1 and Retnla in the above BMDMs induced by the treatment with IL-4 (20 ng/mL) with or without APG-115 were analyzed by RT-qPCR. Duplicated samples were tested. c Western blot analysis of p53, p21, c-Myc and c-Maf total proteins in BMDMs treated with IL-4 (20 ng/mL) with or without APG-115 (1 μM) for 0, 4, or 24 h, or sequentially treated with IL-4 and then APG-115 for 24 h each (24 h + 24 h). d Quantification of C. 0 (black bars), 4 (blue bars), or 24 h (green bars), or sequentially treated with each agent for 24 h (24 h + 24 h, red bars). e naïve BALB/c mice were treated with APG-115 (10 mg/kg, Q2D × 2 doses; n = 5). Two days after the last dose, spleens were collected, dissociated into single-cell suspensions, and stained with macrophage markers for flow cytometry analysis. Macrophages were defined as CD11b + F4/80 hi , and further analyzed for M1 macrophages by expression of MHC-II. Pooled data of percentages of macrophages gated on CD45 + CD3 − live cells ( f ) and percentages of M1 macrophages gated on macrophages ( g ) from five mice were plotted

    Article Snippet: Analyses of T cell activation and proliferation CD4+ T cells were positively selected from mouse spleens using magnetic beads (Miltenyi, catalog #130–049-201) and stimulated with 10 μg/mL plate-bound anti-CD3 (eBioscience, catalog #16–0031-85) and 2 μg/mL anti-CD28 (eBioscience, catalog #16–0281-85) in the presence of 250 nM APG-115 or DMSO for 1 or 2 days.

    Techniques: In Vitro, In Vivo, Activation Assay, Generated, Flow Cytometry, Cytometry, Expressing, Quantitative RT-PCR, Western Blot, Mouse Assay, Staining

    APG-115 increase mouse T cell proliferation and enhances mouse CD4 + T cell activation. a CD4 + T and CD8 + T cells were positively selected from mouse spleens using magnetic beads and then stimulated with indicated concentrations of plate-bound anti-CD3 and 2 μg/mL anti-CD28 in the presence of 250 nM APG-115 or DMSO. After 72 h, relative cell numbers were determined using CellTiter-Glo luminescent cell viability assay (Promega) and normalized to unstimulated cultures treated with DMSO control. * P

    Journal: Journal for Immunotherapy of Cancer

    Article Title: MDM2 inhibitor APG-115 synergizes with PD-1 blockade through enhancing antitumor immunity in the tumor microenvironment

    doi: 10.1186/s40425-019-0750-6

    Figure Lengend Snippet: APG-115 increase mouse T cell proliferation and enhances mouse CD4 + T cell activation. a CD4 + T and CD8 + T cells were positively selected from mouse spleens using magnetic beads and then stimulated with indicated concentrations of plate-bound anti-CD3 and 2 μg/mL anti-CD28 in the presence of 250 nM APG-115 or DMSO. After 72 h, relative cell numbers were determined using CellTiter-Glo luminescent cell viability assay (Promega) and normalized to unstimulated cultures treated with DMSO control. * P

    Article Snippet: Analyses of T cell activation and proliferation CD4+ T cells were positively selected from mouse spleens using magnetic beads (Miltenyi, catalog #130–049-201) and stimulated with 10 μg/mL plate-bound anti-CD3 (eBioscience, catalog #16–0031-85) and 2 μg/mL anti-CD28 (eBioscience, catalog #16–0281-85) in the presence of 250 nM APG-115 or DMSO for 1 or 2 days.

    Techniques: Activation Assay, Magnetic Beads, Cell Viability Assay

    MFG-E8 in ESCs Suppresses T Cell Activation and Proliferation Negatively selected C57BL/6 CD3 + T cells were incubated with different treatment reagents, followed by stimulation with plate-bound anti-CD3/CD28 antibodies. (A) After stimulation, T cells were either untreated (stimulated) or treated with vehicle control (Ct), ESC-derived soluble factors alone (ESC-ext prepared from D3 ESCs derived from 129S2/SvPas mouse blastocysts, 0.23 mg/ml), or in combination with anti-MFG-E8 antibody (2.5 μg/ml) for 24 hr. The cells were then stained for CD4, CD8, and activation markers CD25 and CD69 and subsequently analyzed by flow cytometry. (B) The surface expression of CD25, CD44, and CD69 on CD4 + and CD8 + T cells was analyzed by flow cytometry in untreated control (Ct), stimulated control, recombinant mouse MFG-E8 (rmMFG-E8) alone, rmMFG-E8 with isotype antibodies (iso Ab), rmMFG-E8 with anti-MFG-E8 antibody, differentiated ESC-derived soluble factors (Diff ESC-ext from D3 ESCs), or Diff ESC-ext with rmMFG-E8. The dot plots are all from one experiment in which cells were stained for CD25 and CD69 markers, and three experiments were performed with similar results. Histograms represent the mean fluorescent intensity (MFI) of either CD4 + or CD8 + T cells with CD44 expression for each treatment group. Error bars indicate mean ± SD from three independent experiments; ∗ p

    Journal: Stem Cell Reports

    Article Title: MFG-E8 Is Critical for Embryonic Stem Cell-Mediated T Cell Immunomodulation

    doi: 10.1016/j.stemcr.2015.09.005

    Figure Lengend Snippet: MFG-E8 in ESCs Suppresses T Cell Activation and Proliferation Negatively selected C57BL/6 CD3 + T cells were incubated with different treatment reagents, followed by stimulation with plate-bound anti-CD3/CD28 antibodies. (A) After stimulation, T cells were either untreated (stimulated) or treated with vehicle control (Ct), ESC-derived soluble factors alone (ESC-ext prepared from D3 ESCs derived from 129S2/SvPas mouse blastocysts, 0.23 mg/ml), or in combination with anti-MFG-E8 antibody (2.5 μg/ml) for 24 hr. The cells were then stained for CD4, CD8, and activation markers CD25 and CD69 and subsequently analyzed by flow cytometry. (B) The surface expression of CD25, CD44, and CD69 on CD4 + and CD8 + T cells was analyzed by flow cytometry in untreated control (Ct), stimulated control, recombinant mouse MFG-E8 (rmMFG-E8) alone, rmMFG-E8 with isotype antibodies (iso Ab), rmMFG-E8 with anti-MFG-E8 antibody, differentiated ESC-derived soluble factors (Diff ESC-ext from D3 ESCs), or Diff ESC-ext with rmMFG-E8. The dot plots are all from one experiment in which cells were stained for CD25 and CD69 markers, and three experiments were performed with similar results. Histograms represent the mean fluorescent intensity (MFI) of either CD4 + or CD8 + T cells with CD44 expression for each treatment group. Error bars indicate mean ± SD from three independent experiments; ∗ p

    Article Snippet: Flow Cytometry and Intracellular Staining Fractionated CD3+ T cells were treated with different reagents as described in , followed by stimulation with plate-coated anti-CD3 and anti-CD28 (eBioscience, catalog no. 16-0031-85 and 16-0281-85) for 48 hr.

    Techniques: Activation Assay, Incubation, Derivative Assay, Staining, Flow Cytometry, Cytometry, Expressing, Recombinant

    MFG-E8 in ESCs Suppresses Th1, Th2, and Th17 Subsets while Enhancing the Treg Subpopulation (A) Gene expression of transcriptional factors representing Th1, Th2, and Th17 (qPCR). Purified CD3 + splenic T cells were primed with anti-CD3/CD28 for 3 days and then treated with either D3 ESC-derived soluble factors (D3 ESC-ext, D3 ESC line derived from 129S2/SvPas mouse blastocysts) or rmMFG-E8 in the presence of absence of MFG-E8 blocking antibody, followed by PMA stimulation. Expression levels of mRNA of transcriptional factors ( T-bet , Rorγt , Gata3 , and Foxp3 ) were determined by qPCR. Data represent mean ± SD of at least three independent experiments; ∗ p

    Journal: Stem Cell Reports

    Article Title: MFG-E8 Is Critical for Embryonic Stem Cell-Mediated T Cell Immunomodulation

    doi: 10.1016/j.stemcr.2015.09.005

    Figure Lengend Snippet: MFG-E8 in ESCs Suppresses Th1, Th2, and Th17 Subsets while Enhancing the Treg Subpopulation (A) Gene expression of transcriptional factors representing Th1, Th2, and Th17 (qPCR). Purified CD3 + splenic T cells were primed with anti-CD3/CD28 for 3 days and then treated with either D3 ESC-derived soluble factors (D3 ESC-ext, D3 ESC line derived from 129S2/SvPas mouse blastocysts) or rmMFG-E8 in the presence of absence of MFG-E8 blocking antibody, followed by PMA stimulation. Expression levels of mRNA of transcriptional factors ( T-bet , Rorγt , Gata3 , and Foxp3 ) were determined by qPCR. Data represent mean ± SD of at least three independent experiments; ∗ p

    Article Snippet: Flow Cytometry and Intracellular Staining Fractionated CD3+ T cells were treated with different reagents as described in , followed by stimulation with plate-coated anti-CD3 and anti-CD28 (eBioscience, catalog no. 16-0031-85 and 16-0281-85) for 48 hr.

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Purification, Derivative Assay, Blocking Assay

    MFG-E8 in ESCs Inhibits PKCθ Activation (A) Negatively selected C57BL/6 CD3 + T cells were treated with undifferentiated ESC-derived soluble factors (ESC-ext, prepared from D3 ESCs derived from 129S2/SvPas mouse blastocysts) alone or in combination with either control isotype antibody or anti-MFG-E8 antibody, and stimulated with anti-CD3/CD28 antibodies. For western blot analysis, cell lysates were probed with antibodies to phosphorylated-PKCθ (Ther538), total PKCθ, and α-tubulin (as a loading control). (B) Cells were treated with recombinant mouse MFG-E8 (rmMFG-E8) in the absence or presence of control isotype antibody or anti-MFG-E8 antibody and analyzed by western blot. (C) Cells were treated with soluble factors derived from undifferentiated D3 ESCs (ESC-ext), differentiated D3 ESCs (Diff ESC-ext), or C2C12 myoblast cell line (C2C12 myoblast cell-ext) alone or in combination with rmMFG-E8 and then analyzed by western blot. (D) The αV integrin-blocking antibody (anti-αV Ab) was introduced to restrain the binding of MFG-E8 to its receptor. Cell lysates were then analyzed for levels of PKCθ phosphorylation by densitometry with α-tubulin as a loading control. Bar graphs represent mean ± SD from at least three independent experiments. See also Figure S4 .

    Journal: Stem Cell Reports

    Article Title: MFG-E8 Is Critical for Embryonic Stem Cell-Mediated T Cell Immunomodulation

    doi: 10.1016/j.stemcr.2015.09.005

    Figure Lengend Snippet: MFG-E8 in ESCs Inhibits PKCθ Activation (A) Negatively selected C57BL/6 CD3 + T cells were treated with undifferentiated ESC-derived soluble factors (ESC-ext, prepared from D3 ESCs derived from 129S2/SvPas mouse blastocysts) alone or in combination with either control isotype antibody or anti-MFG-E8 antibody, and stimulated with anti-CD3/CD28 antibodies. For western blot analysis, cell lysates were probed with antibodies to phosphorylated-PKCθ (Ther538), total PKCθ, and α-tubulin (as a loading control). (B) Cells were treated with recombinant mouse MFG-E8 (rmMFG-E8) in the absence or presence of control isotype antibody or anti-MFG-E8 antibody and analyzed by western blot. (C) Cells were treated with soluble factors derived from undifferentiated D3 ESCs (ESC-ext), differentiated D3 ESCs (Diff ESC-ext), or C2C12 myoblast cell line (C2C12 myoblast cell-ext) alone or in combination with rmMFG-E8 and then analyzed by western blot. (D) The αV integrin-blocking antibody (anti-αV Ab) was introduced to restrain the binding of MFG-E8 to its receptor. Cell lysates were then analyzed for levels of PKCθ phosphorylation by densitometry with α-tubulin as a loading control. Bar graphs represent mean ± SD from at least three independent experiments. See also Figure S4 .

    Article Snippet: Flow Cytometry and Intracellular Staining Fractionated CD3+ T cells were treated with different reagents as described in , followed by stimulation with plate-coated anti-CD3 and anti-CD28 (eBioscience, catalog no. 16-0031-85 and 16-0281-85) for 48 hr.

    Techniques: Activation Assay, Derivative Assay, Western Blot, Recombinant, Blocking Assay, Binding Assay

    Impact of Nf1 heterozygosity on T cells proliferation and function. a , b Proliferation of peripheral T cells from Nf1 +/+ and Nf1 +/− mice stimulated by various concentrations of a anti-CD3e antibody alone or with b CD28 costimulation. Proliferation was assessed in biological duplicate and technical triplicate by 3 H-thymidine incorporation during the last 20 h of culture. c Delayed-type hypersensitivity (DTH) assay: On day 0, Nf1 +/+ and Nf1 +/− mice were sensitized on abdominal skin by topical application of oxazolone. On day 6, Nf1 +/+ and Nf1 +/− mice were challenged by topical application on the left and right ear with oxazolone and control solvent, respectively. DTH was assessed daily through by measuring ear thickness (Δ). Experiments were performed in biological triplicates. d Two days after challenge, cellularity of draining lymph nodes (DLNs) of Nf1 +/+ and Nf1 +/− mice were determined. e Spontaneous activation of cells from DLNs (T cells) was measured by 3 H-thymidine incorporation after culturing for 3 days without stimuli. f Fraction of CD4 + cells that are activated. g Fraction of CD8 + cells that are activated. h Fraction of B cells + that are activated. i Fraction of Treg + cells in total CD4 + subset. Panels d – i were performed in biological triplicates. Statistical paired t -test performed; * P

    Journal: Nature Communications

    Article Title: NF1 heterozygosity fosters de novo tumorigenesis but impairs malignant transformation

    doi: 10.1038/s41467-018-07452-y

    Figure Lengend Snippet: Impact of Nf1 heterozygosity on T cells proliferation and function. a , b Proliferation of peripheral T cells from Nf1 +/+ and Nf1 +/− mice stimulated by various concentrations of a anti-CD3e antibody alone or with b CD28 costimulation. Proliferation was assessed in biological duplicate and technical triplicate by 3 H-thymidine incorporation during the last 20 h of culture. c Delayed-type hypersensitivity (DTH) assay: On day 0, Nf1 +/+ and Nf1 +/− mice were sensitized on abdominal skin by topical application of oxazolone. On day 6, Nf1 +/+ and Nf1 +/− mice were challenged by topical application on the left and right ear with oxazolone and control solvent, respectively. DTH was assessed daily through by measuring ear thickness (Δ). Experiments were performed in biological triplicates. d Two days after challenge, cellularity of draining lymph nodes (DLNs) of Nf1 +/+ and Nf1 +/− mice were determined. e Spontaneous activation of cells from DLNs (T cells) was measured by 3 H-thymidine incorporation after culturing for 3 days without stimuli. f Fraction of CD4 + cells that are activated. g Fraction of CD8 + cells that are activated. h Fraction of B cells + that are activated. i Fraction of Treg + cells in total CD4 + subset. Panels d – i were performed in biological triplicates. Statistical paired t -test performed; * P

    Article Snippet: Purified CD3+ T cells (2 × 105 /well) were cultured for 2 days in 96-well plates (in triplicate) precoated with indicated doses of anti-CD3 Ab (eBioscience; cat. no. 16-0031-86) alone or anti-CD28 mAb (eBioscience; cat. no. 16-0281-85).

    Techniques: Mouse Assay, Activation Assay