96 well round bottom plate  (Thermo Fisher)


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    Name:
    96 Well Plate Bulk Round Bottom
    Description:
    Thermo Scientific Abgene polypropylene microplates and deepwell plates offer storage security for assays compound libraries or storing samples for either intermediate or long term use Abgene microtiter plates are manufactured to exacting specifications in our Class 100 000 clean room ISO 9001 conditions using high quality medical grade virgin polypropylene resins which ensure confidence in the quality and performance of the storage plates Designed to ANSI standards these microplates and deepwell plates are compatible with a variety of automated liquid handling for high throughput workflows The polypropylene storage plates are offered with spindle pyramidal U and V bottom wells along with a wide range of volumes to accommodate most applications
    Catalog Number:
    ab0661
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    None
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    Laboratory Plastics and Supplies|Microplates, Dishes and Flasks
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    Lab Supplies Plastics Glassware
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    Structured Review

    Thermo Fisher 96 well round bottom plate
    VDR ligands modulate peptide-specific recall responses from MOG-immunized mice. (a) Compound A inhibits the proliferation of MOG-specific splenocytes. Splenocytes from groups ( n = 5) of MOG-immunized mice were harvested at day 28 and analyzed ex vivo for proliferative responses to MOG at the concentrations indicated. Cells were cultured in triplicate in <t>96-well</t> plates for 60 hours, and proliferation was measured by 3 H-thymidine incorporation during the final 8 hours of culture. Values represent the mean ± SE of triplicate for each peptide concentration. Ovalbumin (OVA) peptide was used as specificity control since the mice were immunized with MOG peptide. Vehicle group consisted of MOG immunized mice treated with vehicle (sesame seed oil). CFA group was mock immunized with CFA only (without MOG peptide) and was not treated with any ligands. (b) Effect of VDR ligands on cytokine elaboration in MOG-immunized animals. For dendritic cell IL-10 production, CD11c + cells were purified on day 28 from splenocytes obtained from MOG-immunized animals. The effect of VDR ligands on IL-10 levels from splenic CD11c + cultures stimulated for 24 hours with 100 ng/mL LPS is shown. VDR ligands decreased IFN- γ elaboration from splenocyte cultures stimulated for 48 hours with 80 μ g/mL MOG peptide. IL-10 and IFN- γ protein levels were measured by ELISA.
    Thermo Scientific Abgene polypropylene microplates and deepwell plates offer storage security for assays compound libraries or storing samples for either intermediate or long term use Abgene microtiter plates are manufactured to exacting specifications in our Class 100 000 clean room ISO 9001 conditions using high quality medical grade virgin polypropylene resins which ensure confidence in the quality and performance of the storage plates Designed to ANSI standards these microplates and deepwell plates are compatible with a variety of automated liquid handling for high throughput workflows The polypropylene storage plates are offered with spindle pyramidal U and V bottom wells along with a wide range of volumes to accommodate most applications
    https://www.bioz.com/result/96 well round bottom plate/product/Thermo Fisher
    Average 99 stars, based on 13 article reviews
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    96 well round bottom plate - by Bioz Stars, 2020-04
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    Images

    1) Product Images from "A Nonsecosteroidal Vitamin D Receptor Modulator Ameliorates Experimental Autoimmune Encephalomyelitis without Causing Hypercalcemia"

    Article Title: A Nonsecosteroidal Vitamin D Receptor Modulator Ameliorates Experimental Autoimmune Encephalomyelitis without Causing Hypercalcemia

    Journal: Autoimmune Diseases

    doi: 10.4061/2011/132958

    VDR ligands modulate peptide-specific recall responses from MOG-immunized mice. (a) Compound A inhibits the proliferation of MOG-specific splenocytes. Splenocytes from groups ( n = 5) of MOG-immunized mice were harvested at day 28 and analyzed ex vivo for proliferative responses to MOG at the concentrations indicated. Cells were cultured in triplicate in 96-well plates for 60 hours, and proliferation was measured by 3 H-thymidine incorporation during the final 8 hours of culture. Values represent the mean ± SE of triplicate for each peptide concentration. Ovalbumin (OVA) peptide was used as specificity control since the mice were immunized with MOG peptide. Vehicle group consisted of MOG immunized mice treated with vehicle (sesame seed oil). CFA group was mock immunized with CFA only (without MOG peptide) and was not treated with any ligands. (b) Effect of VDR ligands on cytokine elaboration in MOG-immunized animals. For dendritic cell IL-10 production, CD11c + cells were purified on day 28 from splenocytes obtained from MOG-immunized animals. The effect of VDR ligands on IL-10 levels from splenic CD11c + cultures stimulated for 24 hours with 100 ng/mL LPS is shown. VDR ligands decreased IFN- γ elaboration from splenocyte cultures stimulated for 48 hours with 80 μ g/mL MOG peptide. IL-10 and IFN- γ protein levels were measured by ELISA.
    Figure Legend Snippet: VDR ligands modulate peptide-specific recall responses from MOG-immunized mice. (a) Compound A inhibits the proliferation of MOG-specific splenocytes. Splenocytes from groups ( n = 5) of MOG-immunized mice were harvested at day 28 and analyzed ex vivo for proliferative responses to MOG at the concentrations indicated. Cells were cultured in triplicate in 96-well plates for 60 hours, and proliferation was measured by 3 H-thymidine incorporation during the final 8 hours of culture. Values represent the mean ± SE of triplicate for each peptide concentration. Ovalbumin (OVA) peptide was used as specificity control since the mice were immunized with MOG peptide. Vehicle group consisted of MOG immunized mice treated with vehicle (sesame seed oil). CFA group was mock immunized with CFA only (without MOG peptide) and was not treated with any ligands. (b) Effect of VDR ligands on cytokine elaboration in MOG-immunized animals. For dendritic cell IL-10 production, CD11c + cells were purified on day 28 from splenocytes obtained from MOG-immunized animals. The effect of VDR ligands on IL-10 levels from splenic CD11c + cultures stimulated for 24 hours with 100 ng/mL LPS is shown. VDR ligands decreased IFN- γ elaboration from splenocyte cultures stimulated for 48 hours with 80 μ g/mL MOG peptide. IL-10 and IFN- γ protein levels were measured by ELISA.

    Techniques Used: Mouse Assay, Ex Vivo, Cell Culture, Concentration Assay, Purification, Enzyme-linked Immunosorbent Assay

    2) Product Images from "Identification and characterization of noncalcemic, tissue-selective, nonsecosteroidal vitamin D receptor modulators"

    Article Title: Identification and characterization of noncalcemic, tissue-selective, nonsecosteroidal vitamin D receptor modulators

    Journal: Journal of Clinical Investigation

    doi: 10.1172/JCI25901

    Nonsecosteroidal VDR ligands are potent agonists in keratinocytes and PBMCs. ( A ) LY2108491 and LY2109866 are potent inhibitors of keratinocyte proliferation. KerTr cells plated in 96-well plates were dosed with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 72 hours at 37°C before BrdU incorporation into DNA was analyzed as a measure of cell proliferation. Results (mean α SEM) of experiments performed in triplicate are shown. ( B ) Nonsecosteroidal VDR ligands are potent inducers of CYP24 gene expression in keratinocytes. TaqMan quantitative RT-PCR (Q-PCR) was performed on total RNA prepared from KerTr cells treated with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. Levels of GAPDH mRNA were measured in all the samples, and the results were normalized and presented as fold induction (α SEM) compared with normalized CYP24 levels in vehicle-treated cells. ( C ) Nonsecosteroidal VDR ligands are efficacious in TPA- and PHA-activated PBMCs. Primary cells isolated from donors were stimulated with TPA (100 ng/ml) and PHA (25 μl/ml) and treated with vehicle or 100 nM each of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. TaqMan Q-PCR was performed on RNA obtained from vehicle-treated or VDR ligand–treated samples, using primer pairs and probes for IL-2, IL-4, IL-10, GATA3, and GAPDH. The amount of IL-2, IL-4, IL-10, and GATA3 transcripts relative to GAPDH transcripts is shown as mean α SEM of quadruplicate experimes.
    Figure Legend Snippet: Nonsecosteroidal VDR ligands are potent agonists in keratinocytes and PBMCs. ( A ) LY2108491 and LY2109866 are potent inhibitors of keratinocyte proliferation. KerTr cells plated in 96-well plates were dosed with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 72 hours at 37°C before BrdU incorporation into DNA was analyzed as a measure of cell proliferation. Results (mean α SEM) of experiments performed in triplicate are shown. ( B ) Nonsecosteroidal VDR ligands are potent inducers of CYP24 gene expression in keratinocytes. TaqMan quantitative RT-PCR (Q-PCR) was performed on total RNA prepared from KerTr cells treated with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. Levels of GAPDH mRNA were measured in all the samples, and the results were normalized and presented as fold induction (α SEM) compared with normalized CYP24 levels in vehicle-treated cells. ( C ) Nonsecosteroidal VDR ligands are efficacious in TPA- and PHA-activated PBMCs. Primary cells isolated from donors were stimulated with TPA (100 ng/ml) and PHA (25 μl/ml) and treated with vehicle or 100 nM each of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. TaqMan Q-PCR was performed on RNA obtained from vehicle-treated or VDR ligand–treated samples, using primer pairs and probes for IL-2, IL-4, IL-10, GATA3, and GAPDH. The amount of IL-2, IL-4, IL-10, and GATA3 transcripts relative to GAPDH transcripts is shown as mean α SEM of quadruplicate experimes.

    Techniques Used: BrdU Incorporation Assay, Expressing, Quantitative RT-PCR, Polymerase Chain Reaction, Isolation

    Related Articles

    Positive Control:

    Article Title: Filarial infection modulates the immune response to Mycobacterium tuberculosis through expansion of CD4+ IL-4 memory T cells
    Article Snippet: Staphylococcus enterotoxin B (SEB) (Toxin Technology Inc., Sarasota, FL) at a final concentration of 1 μg/ml was used as positive control. .. Briefly, cells were plated in RPMI, with 10% Fetal Calf Serum with penicillin/streptomycin (100 U/100 mg/ml), L-glutamine (2 mM) media at a maximum of 2×106 /well, in a volume of 200 ul/well, in a 96-well round bottom plate with BmA, CFP-10 or SEB as well as media alone in the presence of α-CD28/CD49d beads (Invitrogen) at a final concentration of 1 ug/ml, used as co-stimulatory molecules.

    Cytometry:

    Article Title: CD4+ T effector memory cell dysfunction is associated with the accumulation of granulocytic myeloid-derived suppressor cells in glioblastoma patients
    Article Snippet: T-cells were purified using a Pan T-cell isolation kit (MACS, Miltenyi Biotec), resulting in a 95% pure untouched T-cell population as measured by flow cytometry. .. T-cells were labeled with 2 µM CFSE for 5 minutes (Sigma-Aldrich) in phosphate-buffered saline containing 1% fetal calf serum, washed twice, seeded in a 96-well round-bottom plate coated with coated human anti-CD3 (Okt3) mAb (10 µg/mL, eBioscience) at 5 × 104 /well, and co-cultured with MDSC subsets at a cell ratio of 2:1 for 5 days at 37°C in complete medium in the presence of 100 U/mL IL-2 (Peprotech).

    Article Title: Combination of IL-2, rapamycin, DNA methyltransferase and histone deacetylase inhibitors for the expansion of human regulatory T cells
    Article Snippet: Cells were stimulated with 0.5 μg/mL plate-bound anti-CD3 (OKT3 mAb) in 96-well round-bottom plate in RPMI medium supplemented with 10% fetal bovin serum (Bio West), 2mM L-glutamin, 1mM sodium pyruvate,1% non essential amino acid MEM, 100U/mL penicillin, 100 μg/ml streptomycin and amphotericin B (all from Gibco). .. Proliferation of CFSE-labeled cells was assessed by flow cytometry after 84-90 hr of culture.

    Blocking Assay:

    Article Title: Environmental and genetic perturbations reveal different networks of metabolic regulation
    Article Snippet: .. We weighed two replicates of five male or female flies from each block ( ) using a precision balance (Sartorius CP2P) and placed them in 2.0 ml 96-well plates (ABgene, AB-0661). ..

    Incubation:

    Article Title: Increased Frequency of Regulatory T Cells Accompanies Increased Immune Activation in Rectal Mucosae of HIV-Positive Noncontrollers ▿
    Article Snippet: Staining was arrested by the addition of 5 times the volume of ice-cold R15 followed by a 5-min incubation on ice. .. For the suppression assays, wells of a 96-well round-bottom plate were either left uncoated or coated with 0.2 μg/ml anti-CD3, clone OKT3 (eBioscience), for 4 h at 37°C.

    Proliferation Assay:

    Article Title: A Nonsecosteroidal Vitamin D Receptor Modulator Ameliorates Experimental Autoimmune Encephalomyelitis without Causing Hypercalcemia
    Article Snippet: Paragraph title: 2.7. Splenocyte Cell Culture and Proliferation Assay ... Pooled splenocytes of 6 individual mice from the same group were plated in triplicate in 96-well round bottom plate at 2 × 105 cells/well in 200 μ L complete RPMI 1640 medium (Invitrogen) supplemented with 2 mM L-glutamine, 25 mM HEPES, 100 U/mL penicillin, 100 μ g/mL streptomycin, 5.5 × 10−5 M 2-ME, and 5% FCS (all supplements from Invitrogen) containing either 0 and 80 μ g/mL of MOG35-55 (Peptides International) or control OVA323-339 peptide (ISQAVHAAHAEINEAGR, Research Genetics, Inc., Huntsville, AL) and cultured at 37°C, 5% CO2 .

    Flow Cytometry:

    Article Title: CD4+ T effector memory cell dysfunction is associated with the accumulation of granulocytic myeloid-derived suppressor cells in glioblastoma patients
    Article Snippet: T-cells were purified using a Pan T-cell isolation kit (MACS, Miltenyi Biotec), resulting in a 95% pure untouched T-cell population as measured by flow cytometry. .. T-cells were labeled with 2 µM CFSE for 5 minutes (Sigma-Aldrich) in phosphate-buffered saline containing 1% fetal calf serum, washed twice, seeded in a 96-well round-bottom plate coated with coated human anti-CD3 (Okt3) mAb (10 µg/mL, eBioscience) at 5 × 104 /well, and co-cultured with MDSC subsets at a cell ratio of 2:1 for 5 days at 37°C in complete medium in the presence of 100 U/mL IL-2 (Peprotech).

    Article Title: T-cell expression of IL10 is essential for tumor immune surveillance in the small intestine
    Article Snippet: Flow cytometric staining and analysis. .. Cells in suspension were transferred to a 96-well round bottom plate and stained with extracellular antibodies and Live/Dead Violet dye (Invitrogen) for 20 min.

    Article Title: Combination of IL-2, rapamycin, DNA methyltransferase and histone deacetylase inhibitors for the expansion of human regulatory T cells
    Article Snippet: Cells were stimulated with 0.5 μg/mL plate-bound anti-CD3 (OKT3 mAb) in 96-well round-bottom plate in RPMI medium supplemented with 10% fetal bovin serum (Bio West), 2mM L-glutamin, 1mM sodium pyruvate,1% non essential amino acid MEM, 100U/mL penicillin, 100 μg/ml streptomycin and amphotericin B (all from Gibco). .. Proliferation of CFSE-labeled cells was assessed by flow cytometry after 84-90 hr of culture.

    Cell Culture:

    Article Title: T Lymphocyte Inhibition by Tumor-Infiltrating Dendritic Cells Involves Ectonucleotidase CD39 but Not Arginase-1
    Article Snippet: .. T cells, used as responders, were plated in CM at 1 × 105 cells/well in 96-well round bottom plates with anti-CD3/CD28 T-cell expander beads (Invitrogen) and cultured for 4 days with or without DC. ..

    Article Title: Combination of IL-2, rapamycin, DNA methyltransferase and histone deacetylase inhibitors for the expansion of human regulatory T cells
    Article Snippet: Cells were stimulated with 0.5 μg/mL plate-bound anti-CD3 (OKT3 mAb) in 96-well round-bottom plate in RPMI medium supplemented with 10% fetal bovin serum (Bio West), 2mM L-glutamin, 1mM sodium pyruvate,1% non essential amino acid MEM, 100U/mL penicillin, 100 μg/ml streptomycin and amphotericin B (all from Gibco). .. Percent suppression was calculated as follows: [1 - (number of proliferating CFSE diluting responder cells in the presence of suppressor cells at a 1 to 1 ratio / number of proliferating responder cells when cultured alone)] x 100.

    Article Title: Large-scale in vitro expansion of human regulatory T cells with potent xenoantigen-specific suppression
    Article Snippet: .. To obtain high numbers of functionally xenoantigen-specific Treg, 5 × 104 isolated CD4+ CD25+ CD127lo Treg were initially cultured in each well of 96-well round bottom plates with 200 μl RPMI 1640 (Invitrogen, San Diego, CA, USA) containing 10 % human AB serum (Invitrogen), 2 mM glutamine, 25 mM HEPES, 50 U/ml penicillin, 50 μg/ml streptomycin, 50 μM 2-mercaptoethanol (Sigma, St. Louis, MO, USA), and 100 nM rapamycin (Sigma) at 37 °C and 5 % CO2 , in the presence of 400 U/ml IL-2 (Chiron, Emeryville, CA, USA) and T cell expander beads (CD3/CD28 Dynabeads; Invitrogen Dynal) at a ratio of 4 beads per cell for 7 days as polyclonal stimulation. .. The polyclonally stimulated Treg were then expanded with three subsequent cycles of xenoantigen stimulation (7 days per cycle) using irradiated (30 Gy) pig PBMC at a 4:1 ratio of xenoPBMC:Treg in the presence of 400 U/ml IL-2 and T cell expander beads at a 1:1 ratio of beads:Treg (Fig. ).

    Article Title: A Nonsecosteroidal Vitamin D Receptor Modulator Ameliorates Experimental Autoimmune Encephalomyelitis without Causing Hypercalcemia
    Article Snippet: .. Pooled splenocytes of 6 individual mice from the same group were plated in triplicate in 96-well round bottom plate at 2 × 105 cells/well in 200 μ L complete RPMI 1640 medium (Invitrogen) supplemented with 2 mM L-glutamine, 25 mM HEPES, 100 U/mL penicillin, 100 μ g/mL streptomycin, 5.5 × 10−5 M 2-ME, and 5% FCS (all supplements from Invitrogen) containing either 0 and 80 μ g/mL of MOG35-55 (Peptides International) or control OVA323-339 peptide (ISQAVHAAHAEINEAGR, Research Genetics, Inc., Huntsville, AL) and cultured at 37°C, 5% CO2 . .. Proliferation was measured by incorporation of [3 H]-methylthymidine (1 μ Ci/well, ICN Radiochemicals, Irvine, CA) during the last 8 hr of culture using a filtermate harvester (Packard Instrument Co., Downers Grove, IL) and a 1450 microbeta liquid scintillation counter (Pharmacia Biotech AB).

    Article Title: Identification and characterization of noncalcemic, tissue-selective, nonsecosteroidal vitamin D receptor modulators
    Article Snippet: .. Pooled splenocytes of 5 individual mice from the same group were plated in triplicate in a 96-well round-bottom plate at 2 × 105 cells per well in 200 μl complete RPMI 1640 medium (Invitrogen Corp.) supplemented with 2 mM l-glutamine, 25 mM HEPES, 100 U/ml penicillin, 100 μg/ml streptomycin, 5.5 × 10–5 M 2-mercaptoethanol, and 5% FCS (all supplements from Invitrogen Corp.) containing 10 μg/ml MOG35–55 (Peptides International Inc.) or medium, and cultured at 37°C, 5% CO2 for 72 hours. .. Cytokine levels produced from cultured splenocytes were analyzed with R & D Systems MAP kit (R & D Systems) (LUM000) with IL-2 (LUM402) and IFN-γ (LUM485) beads.

    Injection:

    Article Title: T-cell expression of IL10 is essential for tumor immune surveillance in the small intestine
    Article Snippet: Lin-depleted bone marrow cells were injected into the retro-orbital (RO) sinus. .. Cells in suspension were transferred to a 96-well round bottom plate and stained with extracellular antibodies and Live/Dead Violet dye (Invitrogen) for 20 min.

    Recombinant:

    Article Title: Thrombin generation measurement using the ST Genesia Thrombin Generation System in a cohort of healthy adults: Normal values and variability, et al. Thrombin generation measurement using the ST Genesia Thrombin Generation System in a cohort of healthy adults: Normal values and variability
    Article Snippet: .. For the second setting, 74 μL PFP was added to 20 μL of a mixture of 5 pmol/L TF and 4 μmol/L phospolipids (PPP Reagent) and 6 μL of recombinant human TM (4 nmol/L, Sekisui Alveo AG, Lucern, Switzerland) or HN‐buffer (HEPES 20 mmol/L, NaCl 140 mmol/L, pH 7.4 + 5 mg/mL BSA), in a 96‐well round‐bottom microtiter plate (Immulon2HB). .. The reaction was initiated with 20 μl of a mixture of fluorogenic substrate and CaCl2 (Fluobuffer, Stago) and fluorescence measured using a Fluoroskan Ascent reader (Thermo Labsystems).

    Magnetic Cell Separation:

    Article Title: CD4+ T effector memory cell dysfunction is associated with the accumulation of granulocytic myeloid-derived suppressor cells in glioblastoma patients
    Article Snippet: T-cells were purified using a Pan T-cell isolation kit (MACS, Miltenyi Biotec), resulting in a 95% pure untouched T-cell population as measured by flow cytometry. .. T-cells were labeled with 2 µM CFSE for 5 minutes (Sigma-Aldrich) in phosphate-buffered saline containing 1% fetal calf serum, washed twice, seeded in a 96-well round-bottom plate coated with coated human anti-CD3 (Okt3) mAb (10 µg/mL, eBioscience) at 5 × 104 /well, and co-cultured with MDSC subsets at a cell ratio of 2:1 for 5 days at 37°C in complete medium in the presence of 100 U/mL IL-2 (Peprotech).

    Fluorescence:

    Article Title: CD4+ T effector memory cell dysfunction is associated with the accumulation of granulocytic myeloid-derived suppressor cells in glioblastoma patients
    Article Snippet: T-cells were labeled with 2 µM CFSE for 5 minutes (Sigma-Aldrich) in phosphate-buffered saline containing 1% fetal calf serum, washed twice, seeded in a 96-well round-bottom plate coated with coated human anti-CD3 (Okt3) mAb (10 µg/mL, eBioscience) at 5 × 104 /well, and co-cultured with MDSC subsets at a cell ratio of 2:1 for 5 days at 37°C in complete medium in the presence of 100 U/mL IL-2 (Peprotech). .. T-cell proliferation was monitored by flow cytometric analysis of CFSE fluorescence intensity after staining with PC5.5-labeled anti-CD3 mAb.

    Magnetic Beads:

    Article Title: Increased Frequency of Regulatory T Cells Accompanies Increased Immune Activation in Rectal Mucosae of HIV-Positive Noncontrollers ▿
    Article Snippet: Briefly, after incubation with the EasySep human CD4 enrichment cocktail, magnetic beads (“D particles”) were used to negatively select for CD4+ T cells. .. For the suppression assays, wells of a 96-well round-bottom plate were either left uncoated or coated with 0.2 μg/ml anti-CD3, clone OKT3 (eBioscience), for 4 h at 37°C.

    Isolation:

    Article Title: CD4+ T effector memory cell dysfunction is associated with the accumulation of granulocytic myeloid-derived suppressor cells in glioblastoma patients
    Article Snippet: Paragraph title: Isolation of MDSC Subsets and T-Cell Assays ... T-cells were labeled with 2 µM CFSE for 5 minutes (Sigma-Aldrich) in phosphate-buffered saline containing 1% fetal calf serum, washed twice, seeded in a 96-well round-bottom plate coated with coated human anti-CD3 (Okt3) mAb (10 µg/mL, eBioscience) at 5 × 104 /well, and co-cultured with MDSC subsets at a cell ratio of 2:1 for 5 days at 37°C in complete medium in the presence of 100 U/mL IL-2 (Peprotech).

    Article Title: Large-scale in vitro expansion of human regulatory T cells with potent xenoantigen-specific suppression
    Article Snippet: .. To obtain high numbers of functionally xenoantigen-specific Treg, 5 × 104 isolated CD4+ CD25+ CD127lo Treg were initially cultured in each well of 96-well round bottom plates with 200 μl RPMI 1640 (Invitrogen, San Diego, CA, USA) containing 10 % human AB serum (Invitrogen), 2 mM glutamine, 25 mM HEPES, 50 U/ml penicillin, 50 μg/ml streptomycin, 50 μM 2-mercaptoethanol (Sigma, St. Louis, MO, USA), and 100 nM rapamycin (Sigma) at 37 °C and 5 % CO2 , in the presence of 400 U/ml IL-2 (Chiron, Emeryville, CA, USA) and T cell expander beads (CD3/CD28 Dynabeads; Invitrogen Dynal) at a ratio of 4 beads per cell for 7 days as polyclonal stimulation. .. The polyclonally stimulated Treg were then expanded with three subsequent cycles of xenoantigen stimulation (7 days per cycle) using irradiated (30 Gy) pig PBMC at a 4:1 ratio of xenoPBMC:Treg in the presence of 400 U/ml IL-2 and T cell expander beads at a 1:1 ratio of beads:Treg (Fig. ).

    Article Title: A Nonsecosteroidal Vitamin D Receptor Modulator Ameliorates Experimental Autoimmune Encephalomyelitis without Causing Hypercalcemia
    Article Snippet: Splenocyte Cell Culture and Proliferation Assay Splenocyte cell suspensions were isolated from MOG35-55 -immunized mice at day 28 by homogenizing spleens between frosted glass slides (Fisher, Pittsburgh, PA) and removing RBC with ACK lysing buffer (BioWhittaker, Walkersville, MD). .. Pooled splenocytes of 6 individual mice from the same group were plated in triplicate in 96-well round bottom plate at 2 × 105 cells/well in 200 μ L complete RPMI 1640 medium (Invitrogen) supplemented with 2 mM L-glutamine, 25 mM HEPES, 100 U/mL penicillin, 100 μ g/mL streptomycin, 5.5 × 10−5 M 2-ME, and 5% FCS (all supplements from Invitrogen) containing either 0 and 80 μ g/mL of MOG35-55 (Peptides International) or control OVA323-339 peptide (ISQAVHAAHAEINEAGR, Research Genetics, Inc., Huntsville, AL) and cultured at 37°C, 5% CO2 .

    Article Title: Identification and characterization of noncalcemic, tissue-selective, nonsecosteroidal vitamin D receptor modulators
    Article Snippet: Ten days later, splenocytes were isolated by homogenizing spleens between frosted glass slides (Fisher Scientific International), and rbcs were removed with ACK lysing buffer (BioWhittaker Inc.). .. Pooled splenocytes of 5 individual mice from the same group were plated in triplicate in a 96-well round-bottom plate at 2 × 105 cells per well in 200 μl complete RPMI 1640 medium (Invitrogen Corp.) supplemented with 2 mM l-glutamine, 25 mM HEPES, 100 U/ml penicillin, 100 μg/ml streptomycin, 5.5 × 10–5 M 2-mercaptoethanol, and 5% FCS (all supplements from Invitrogen Corp.) containing 10 μg/ml MOG35–55 (Peptides International Inc.) or medium, and cultured at 37°C, 5% CO2 for 72 hours.

    Labeling:

    Article Title: CD4+ T effector memory cell dysfunction is associated with the accumulation of granulocytic myeloid-derived suppressor cells in glioblastoma patients
    Article Snippet: .. T-cells were labeled with 2 µM CFSE for 5 minutes (Sigma-Aldrich) in phosphate-buffered saline containing 1% fetal calf serum, washed twice, seeded in a 96-well round-bottom plate coated with coated human anti-CD3 (Okt3) mAb (10 µg/mL, eBioscience) at 5 × 104 /well, and co-cultured with MDSC subsets at a cell ratio of 2:1 for 5 days at 37°C in complete medium in the presence of 100 U/mL IL-2 (Peprotech). .. T-cell proliferation was monitored by flow cytometric analysis of CFSE fluorescence intensity after staining with PC5.5-labeled anti-CD3 mAb.

    Mouse Assay:

    Article Title: T Lymphocyte Inhibition by Tumor-Infiltrating Dendritic Cells Involves Ectonucleotidase CD39 but Not Arginase-1
    Article Snippet: T-Cell Proliferation Assays Splenocytes from naïve BALB/c or C57BL/6 mice were enriched for T lymphocytes using nylon wool columns. .. T cells, used as responders, were plated in CM at 1 × 105 cells/well in 96-well round bottom plates with anti-CD3/CD28 T-cell expander beads (Invitrogen) and cultured for 4 days with or without DC.

    Article Title: A Nonsecosteroidal Vitamin D Receptor Modulator Ameliorates Experimental Autoimmune Encephalomyelitis without Causing Hypercalcemia
    Article Snippet: .. Pooled splenocytes of 6 individual mice from the same group were plated in triplicate in 96-well round bottom plate at 2 × 105 cells/well in 200 μ L complete RPMI 1640 medium (Invitrogen) supplemented with 2 mM L-glutamine, 25 mM HEPES, 100 U/mL penicillin, 100 μ g/mL streptomycin, 5.5 × 10−5 M 2-ME, and 5% FCS (all supplements from Invitrogen) containing either 0 and 80 μ g/mL of MOG35-55 (Peptides International) or control OVA323-339 peptide (ISQAVHAAHAEINEAGR, Research Genetics, Inc., Huntsville, AL) and cultured at 37°C, 5% CO2 . .. Proliferation was measured by incorporation of [3 H]-methylthymidine (1 μ Ci/well, ICN Radiochemicals, Irvine, CA) during the last 8 hr of culture using a filtermate harvester (Packard Instrument Co., Downers Grove, IL) and a 1450 microbeta liquid scintillation counter (Pharmacia Biotech AB).

    Article Title: Identification and characterization of noncalcemic, tissue-selective, nonsecosteroidal vitamin D receptor modulators
    Article Snippet: .. Pooled splenocytes of 5 individual mice from the same group were plated in triplicate in a 96-well round-bottom plate at 2 × 105 cells per well in 200 μl complete RPMI 1640 medium (Invitrogen Corp.) supplemented with 2 mM l-glutamine, 25 mM HEPES, 100 U/ml penicillin, 100 μg/ml streptomycin, 5.5 × 10–5 M 2-mercaptoethanol, and 5% FCS (all supplements from Invitrogen Corp.) containing 10 μg/ml MOG35–55 (Peptides International Inc.) or medium, and cultured at 37°C, 5% CO2 for 72 hours. .. Cytokine levels produced from cultured splenocytes were analyzed with R & D Systems MAP kit (R & D Systems) (LUM000) with IL-2 (LUM402) and IFN-γ (LUM485) beads.

    FACS:

    Article Title: CD4+ T effector memory cell dysfunction is associated with the accumulation of granulocytic myeloid-derived suppressor cells in glioblastoma patients
    Article Snippet: Freshly prepared PBMCs and tumor cell suspensions were labeled with CD45 Krome Orange, CD11b-PC5.5, CD14-FITC, CD15-PB mAbs (Beckman Coulter) and sorted with a FACS Aria III (Becton Dickinson) to isolate different MDSC subsets. .. T-cells were labeled with 2 µM CFSE for 5 minutes (Sigma-Aldrich) in phosphate-buffered saline containing 1% fetal calf serum, washed twice, seeded in a 96-well round-bottom plate coated with coated human anti-CD3 (Okt3) mAb (10 µg/mL, eBioscience) at 5 × 104 /well, and co-cultured with MDSC subsets at a cell ratio of 2:1 for 5 days at 37°C in complete medium in the presence of 100 U/mL IL-2 (Peprotech).

    Purification:

    Article Title: CD4+ T effector memory cell dysfunction is associated with the accumulation of granulocytic myeloid-derived suppressor cells in glioblastoma patients
    Article Snippet: T-cells were purified using a Pan T-cell isolation kit (MACS, Miltenyi Biotec), resulting in a 95% pure untouched T-cell population as measured by flow cytometry. .. T-cells were labeled with 2 µM CFSE for 5 minutes (Sigma-Aldrich) in phosphate-buffered saline containing 1% fetal calf serum, washed twice, seeded in a 96-well round-bottom plate coated with coated human anti-CD3 (Okt3) mAb (10 µg/mL, eBioscience) at 5 × 104 /well, and co-cultured with MDSC subsets at a cell ratio of 2:1 for 5 days at 37°C in complete medium in the presence of 100 U/mL IL-2 (Peprotech).

    Article Title: A Nonsecosteroidal Vitamin D Receptor Modulator Ameliorates Experimental Autoimmune Encephalomyelitis without Causing Hypercalcemia
    Article Snippet: Pooled splenocytes of 6 individual mice from the same group were plated in triplicate in 96-well round bottom plate at 2 × 105 cells/well in 200 μ L complete RPMI 1640 medium (Invitrogen) supplemented with 2 mM L-glutamine, 25 mM HEPES, 100 U/mL penicillin, 100 μ g/mL streptomycin, 5.5 × 10−5 M 2-ME, and 5% FCS (all supplements from Invitrogen) containing either 0 and 80 μ g/mL of MOG35-55 (Peptides International) or control OVA323-339 peptide (ISQAVHAAHAEINEAGR, Research Genetics, Inc., Huntsville, AL) and cultured at 37°C, 5% CO2 . .. Cytokine levels produced by cultured splenocytes or purified CD11c+ dendritic cells from splenocytes (Miltenyi Biotech.) from MOG35-55 -immunized mice were analyzed by removing 100 μ L of cell culture supernatant per well after 60 h of culture as described above.

    Article Title: Increased Frequency of Regulatory T Cells Accompanies Increased Immune Activation in Rectal Mucosae of HIV-Positive Noncontrollers ▿
    Article Snippet: Paragraph title: Purification of T cell subsets and suppression assays. ... For the suppression assays, wells of a 96-well round-bottom plate were either left uncoated or coated with 0.2 μg/ml anti-CD3, clone OKT3 (eBioscience), for 4 h at 37°C.

    Software:

    Article Title: T-cell expression of IL10 is essential for tumor immune surveillance in the small intestine
    Article Snippet: Cells in suspension were transferred to a 96-well round bottom plate and stained with extracellular antibodies and Live/Dead Violet dye (Invitrogen) for 20 min. .. Data were acquired on a FACSCanto II instrument, and analyzed using FlowJo software (TreeStar).

    Irradiation:

    Article Title: Combination of IL-2, rapamycin, DNA methyltransferase and histone deacetylase inhibitors for the expansion of human regulatory T cells
    Article Snippet: 1 × 104 CFSE (1μM Invitrogen)-labeled responder CD25− CD45RA+ CD4+ T cells were cocultured with 1 × 104 unlabeled cells assessed for their suppressive capacity and 1 × 105 irradiated autologous accessory cells containing B cells and monocytes. .. Cells were stimulated with 0.5 μg/mL plate-bound anti-CD3 (OKT3 mAb) in 96-well round-bottom plate in RPMI medium supplemented with 10% fetal bovin serum (Bio West), 2mM L-glutamin, 1mM sodium pyruvate,1% non essential amino acid MEM, 100U/mL penicillin, 100 μg/ml streptomycin and amphotericin B (all from Gibco).

    Article Title: Large-scale in vitro expansion of human regulatory T cells with potent xenoantigen-specific suppression
    Article Snippet: To obtain high numbers of functionally xenoantigen-specific Treg, 5 × 104 isolated CD4+ CD25+ CD127lo Treg were initially cultured in each well of 96-well round bottom plates with 200 μl RPMI 1640 (Invitrogen, San Diego, CA, USA) containing 10 % human AB serum (Invitrogen), 2 mM glutamine, 25 mM HEPES, 50 U/ml penicillin, 50 μg/ml streptomycin, 50 μM 2-mercaptoethanol (Sigma, St. Louis, MO, USA), and 100 nM rapamycin (Sigma) at 37 °C and 5 % CO2 , in the presence of 400 U/ml IL-2 (Chiron, Emeryville, CA, USA) and T cell expander beads (CD3/CD28 Dynabeads; Invitrogen Dynal) at a ratio of 4 beads per cell for 7 days as polyclonal stimulation. .. The polyclonally stimulated Treg were then expanded with three subsequent cycles of xenoantigen stimulation (7 days per cycle) using irradiated (30 Gy) pig PBMC at a 4:1 ratio of xenoPBMC:Treg in the presence of 400 U/ml IL-2 and T cell expander beads at a 1:1 ratio of beads:Treg (Fig. ).

    Article Title: Increased Frequency of Regulatory T Cells Accompanies Increased Immune Activation in Rectal Mucosae of HIV-Positive Noncontrollers ▿
    Article Snippet: For the suppression assays, wells of a 96-well round-bottom plate were either left uncoated or coated with 0.2 μg/ml anti-CD3, clone OKT3 (eBioscience), for 4 h at 37°C. .. Autologous PBMC were irradiated at 4,000 rads; 1 × 105 irradiated feeder cells were added to the culture wells along with 1 × 104 to 11 × 104 CFSE-labeled responder cells and the indicated number of CD4+ CD25+ Treg in R15 supplemented with 10 mM HEPES, 1 mM sodium pyruvate, and 0.1 mM nonessential amino acids.

    Selection:

    Article Title: Increased Frequency of Regulatory T Cells Accompanies Increased Immune Activation in Rectal Mucosae of HIV-Positive Noncontrollers ▿
    Article Snippet: The CD4-enriched cells were then incubated with EasySep human CD25 selection cocktail and magnetic nanoparticles to positively select for the CD4+ CD25-bright population. .. For the suppression assays, wells of a 96-well round-bottom plate were either left uncoated or coated with 0.2 μg/ml anti-CD3, clone OKT3 (eBioscience), for 4 h at 37°C.

    In Vitro:

    Article Title: Combination of IL-2, rapamycin, DNA methyltransferase and histone deacetylase inhibitors for the expansion of human regulatory T cells
    Article Snippet: Paragraph title: In vitro suppressive assays ... Cells were stimulated with 0.5 μg/mL plate-bound anti-CD3 (OKT3 mAb) in 96-well round-bottom plate in RPMI medium supplemented with 10% fetal bovin serum (Bio West), 2mM L-glutamin, 1mM sodium pyruvate,1% non essential amino acid MEM, 100U/mL penicillin, 100 μg/ml streptomycin and amphotericin B (all from Gibco).

    Article Title: Filarial infection modulates the immune response to Mycobacterium tuberculosis through expansion of CD4+ IL-4 memory T cells
    Article Snippet: Paragraph title: Antigens and In Vitro Culture ... Briefly, cells were plated in RPMI, with 10% Fetal Calf Serum with penicillin/streptomycin (100 U/100 mg/ml), L-glutamine (2 mM) media at a maximum of 2×106 /well, in a volume of 200 ul/well, in a 96-well round bottom plate with BmA, CFP-10 or SEB as well as media alone in the presence of α-CD28/CD49d beads (Invitrogen) at a final concentration of 1 ug/ml, used as co-stimulatory molecules.

    Produced:

    Article Title: A Nonsecosteroidal Vitamin D Receptor Modulator Ameliorates Experimental Autoimmune Encephalomyelitis without Causing Hypercalcemia
    Article Snippet: Pooled splenocytes of 6 individual mice from the same group were plated in triplicate in 96-well round bottom plate at 2 × 105 cells/well in 200 μ L complete RPMI 1640 medium (Invitrogen) supplemented with 2 mM L-glutamine, 25 mM HEPES, 100 U/mL penicillin, 100 μ g/mL streptomycin, 5.5 × 10−5 M 2-ME, and 5% FCS (all supplements from Invitrogen) containing either 0 and 80 μ g/mL of MOG35-55 (Peptides International) or control OVA323-339 peptide (ISQAVHAAHAEINEAGR, Research Genetics, Inc., Huntsville, AL) and cultured at 37°C, 5% CO2 . .. Cytokine levels produced by cultured splenocytes or purified CD11c+ dendritic cells from splenocytes (Miltenyi Biotech.) from MOG35-55 -immunized mice were analyzed by removing 100 μ L of cell culture supernatant per well after 60 h of culture as described above.

    Article Title: Identification and characterization of noncalcemic, tissue-selective, nonsecosteroidal vitamin D receptor modulators
    Article Snippet: Pooled splenocytes of 5 individual mice from the same group were plated in triplicate in a 96-well round-bottom plate at 2 × 105 cells per well in 200 μl complete RPMI 1640 medium (Invitrogen Corp.) supplemented with 2 mM l-glutamine, 25 mM HEPES, 100 U/ml penicillin, 100 μg/ml streptomycin, 5.5 × 10–5 M 2-mercaptoethanol, and 5% FCS (all supplements from Invitrogen Corp.) containing 10 μg/ml MOG35–55 (Peptides International Inc.) or medium, and cultured at 37°C, 5% CO2 for 72 hours. .. Cytokine levels produced from cultured splenocytes were analyzed with R & D Systems MAP kit (R & D Systems) (LUM000) with IL-2 (LUM402) and IFN-γ (LUM485) beads.

    Concentration Assay:

    Article Title: Drosophila Lung Cancer Models Identify Trametinib Plus Statin as Candidate Therapeutic
    Article Snippet: The Selleck FDA-approved library (catalog #L1300) was dissolved in DMSO buffer to a 100 mM concentration. .. A PerkinElmer JANUS automated liquid handler was used to create 1.2ml 96 well plates (Abgene catalog #Ab-1068) containing 300μl fly food and 0.3μl drug per well.

    Article Title: Filarial infection modulates the immune response to Mycobacterium tuberculosis through expansion of CD4+ IL-4 memory T cells
    Article Snippet: .. Briefly, cells were plated in RPMI, with 10% Fetal Calf Serum with penicillin/streptomycin (100 U/100 mg/ml), L-glutamine (2 mM) media at a maximum of 2×106 /well, in a volume of 200 ul/well, in a 96-well round bottom plate with BmA, CFP-10 or SEB as well as media alone in the presence of α-CD28/CD49d beads (Invitrogen) at a final concentration of 1 ug/ml, used as co-stimulatory molecules. ..

    Staining:

    Article Title: CD4+ T effector memory cell dysfunction is associated with the accumulation of granulocytic myeloid-derived suppressor cells in glioblastoma patients
    Article Snippet: Cytospins were prepared from each MDSC subset, and cells were stained with hematoxylin and eosin (H & E) to assess their morphology and purity. .. T-cells were labeled with 2 µM CFSE for 5 minutes (Sigma-Aldrich) in phosphate-buffered saline containing 1% fetal calf serum, washed twice, seeded in a 96-well round-bottom plate coated with coated human anti-CD3 (Okt3) mAb (10 µg/mL, eBioscience) at 5 × 104 /well, and co-cultured with MDSC subsets at a cell ratio of 2:1 for 5 days at 37°C in complete medium in the presence of 100 U/mL IL-2 (Peprotech).

    Article Title: T-cell expression of IL10 is essential for tumor immune surveillance in the small intestine
    Article Snippet: .. Cells in suspension were transferred to a 96-well round bottom plate and stained with extracellular antibodies and Live/Dead Violet dye (Invitrogen) for 20 min. .. Cells were then washed and fixed with 1% PFA (10 min), permeabilized (0.3% saponin), and stained with intracellular antibodies (30 min).

    Article Title: Increased Frequency of Regulatory T Cells Accompanies Increased Immune Activation in Rectal Mucosae of HIV-Positive Noncontrollers ▿
    Article Snippet: Staining was arrested by the addition of 5 times the volume of ice-cold R15 followed by a 5-min incubation on ice. .. For the suppression assays, wells of a 96-well round-bottom plate were either left uncoated or coated with 0.2 μg/ml anti-CD3, clone OKT3 (eBioscience), for 4 h at 37°C.

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    Thermo Fisher rpmi 1640 medium
    Infected IL-17A −/− mice show dominant filarial-specific IFN-γ responses. On day 28 p.i., draining mLN cells (5 × 10 5 cells/well) from individual mice were plated in <t>RPMI</t> 1640 medium with supplements and left either alone (Cont.) or stimulated with LsAg (50 μg/ml) or αCD3/αCD28 (5/1.25 μg/ml) in triplicates. After 72 h, the culture supernatant was removed and screened for the presence of IFN-γ ( a ), IL-4 ( b ), IL-6 ( c ) and IL-21 ( d ) by ELISA. Graphs show cytokine responses and values are expressed as mean ± SEM from each mouse from three independent infection experiments ( n = 10 IL-17A −/− and n = 13 WT mice). Statistical significances between the indicated groups were obtained using the Mann-Whitney U tests. Asterisks denote significant differences between the groups indicated by the brackets (* p
    Rpmi 1640 Medium, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 16244 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rpmi 1640 medium/product/Thermo Fisher
    Average 99 stars, based on 16244 article reviews
    Price from $9.99 to $1999.99
    rpmi 1640 medium - by Bioz Stars, 2020-04
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    80
    Thermo Fisher nunclon delta surface 96 well round bottom plates
    Infected IL-17A −/− mice show dominant filarial-specific IFN-γ responses. On day 28 p.i., draining mLN cells (5 × 10 5 cells/well) from individual mice were plated in <t>RPMI</t> 1640 medium with supplements and left either alone (Cont.) or stimulated with LsAg (50 μg/ml) or αCD3/αCD28 (5/1.25 μg/ml) in triplicates. After 72 h, the culture supernatant was removed and screened for the presence of IFN-γ ( a ), IL-4 ( b ), IL-6 ( c ) and IL-21 ( d ) by ELISA. Graphs show cytokine responses and values are expressed as mean ± SEM from each mouse from three independent infection experiments ( n = 10 IL-17A −/− and n = 13 WT mice). Statistical significances between the indicated groups were obtained using the Mann-Whitney U tests. Asterisks denote significant differences between the groups indicated by the brackets (* p
    Nunclon Delta Surface 96 Well Round Bottom Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nunclon delta surface 96 well round bottom plates/product/Thermo Fisher
    Average 80 stars, based on 1 article reviews
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    nunclon delta surface 96 well round bottom plates - by Bioz Stars, 2020-04
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    95
    Thermo Fisher nunc 96 well round bottom black polypropylene microtiter plates
    Infected IL-17A −/− mice show dominant filarial-specific IFN-γ responses. On day 28 p.i., draining mLN cells (5 × 10 5 cells/well) from individual mice were plated in <t>RPMI</t> 1640 medium with supplements and left either alone (Cont.) or stimulated with LsAg (50 μg/ml) or αCD3/αCD28 (5/1.25 μg/ml) in triplicates. After 72 h, the culture supernatant was removed and screened for the presence of IFN-γ ( a ), IL-4 ( b ), IL-6 ( c ) and IL-21 ( d ) by ELISA. Graphs show cytokine responses and values are expressed as mean ± SEM from each mouse from three independent infection experiments ( n = 10 IL-17A −/− and n = 13 WT mice). Statistical significances between the indicated groups were obtained using the Mann-Whitney U tests. Asterisks denote significant differences between the groups indicated by the brackets (* p
    Nunc 96 Well Round Bottom Black Polypropylene Microtiter Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nunc 96 well round bottom black polypropylene microtiter plates/product/Thermo Fisher
    Average 95 stars, based on 3 article reviews
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    nunc 96 well round bottom black polypropylene microtiter plates - by Bioz Stars, 2020-04
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    Infected IL-17A −/− mice show dominant filarial-specific IFN-γ responses. On day 28 p.i., draining mLN cells (5 × 10 5 cells/well) from individual mice were plated in RPMI 1640 medium with supplements and left either alone (Cont.) or stimulated with LsAg (50 μg/ml) or αCD3/αCD28 (5/1.25 μg/ml) in triplicates. After 72 h, the culture supernatant was removed and screened for the presence of IFN-γ ( a ), IL-4 ( b ), IL-6 ( c ) and IL-21 ( d ) by ELISA. Graphs show cytokine responses and values are expressed as mean ± SEM from each mouse from three independent infection experiments ( n = 10 IL-17A −/− and n = 13 WT mice). Statistical significances between the indicated groups were obtained using the Mann-Whitney U tests. Asterisks denote significant differences between the groups indicated by the brackets (* p

    Journal: Parasitology Research

    Article Title: Absence of IL-17A in Litomosoides sigmodontis-infected mice influences worm development and drives elevated filarial-specific IFN-γ

    doi: 10.1007/s00436-018-5959-7

    Figure Lengend Snippet: Infected IL-17A −/− mice show dominant filarial-specific IFN-γ responses. On day 28 p.i., draining mLN cells (5 × 10 5 cells/well) from individual mice were plated in RPMI 1640 medium with supplements and left either alone (Cont.) or stimulated with LsAg (50 μg/ml) or αCD3/αCD28 (5/1.25 μg/ml) in triplicates. After 72 h, the culture supernatant was removed and screened for the presence of IFN-γ ( a ), IL-4 ( b ), IL-6 ( c ) and IL-21 ( d ) by ELISA. Graphs show cytokine responses and values are expressed as mean ± SEM from each mouse from three independent infection experiments ( n = 10 IL-17A −/− and n = 13 WT mice). Statistical significances between the indicated groups were obtained using the Mann-Whitney U tests. Asterisks denote significant differences between the groups indicated by the brackets (* p

    Article Snippet: For bulk cell assays, 5 × 105 erythrocyte-depleted mLN cells from individual mice were plated in 96-well round-bottomed plates in RPMI 1640 medium containing 10% FCS, 1% Pen/Strep, 1% L-glutamine and 0.1% gentamycin (Thermo Fisher Scientific).

    Techniques: Infection, Mouse Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

    Infected IL-17A −/− mice show dominant filarial-specific IFN-γ responses. On day 28 p.i., draining mLN cells (5 × 10 5 cells/well) from individual mice were plated in RPMI 1640 medium with supplements and left either alone (Cont.) or stimulated with LsAg (50 μg/ml) or αCD3/αCD28 (5/1.25 μg/ml) in triplicates. After 72 h, the culture supernatant was removed and screened for the presence of IFN-γ ( a ), IL-4 ( b ), IL-6 ( c ) and IL-21 ( d ) by ELISA. Graphs show cytokine responses and values are expressed as mean ± SEM from each mouse from three independent infection experiments ( n = 10 IL-17A −/− and n = 13 WT mice). Statistical significances between the indicated groups were obtained using the Mann-Whitney U tests. Asterisks denote significant differences between the groups indicated by the brackets (* p

    Journal: Parasitology Research

    Article Title: Absence of IL-17A in Litomosoides sigmodontis-infected mice influences worm development and drives elevated filarial-specific IFN-γ

    doi: 10.1007/s00436-018-5959-7

    Figure Lengend Snippet: Infected IL-17A −/− mice show dominant filarial-specific IFN-γ responses. On day 28 p.i., draining mLN cells (5 × 10 5 cells/well) from individual mice were plated in RPMI 1640 medium with supplements and left either alone (Cont.) or stimulated with LsAg (50 μg/ml) or αCD3/αCD28 (5/1.25 μg/ml) in triplicates. After 72 h, the culture supernatant was removed and screened for the presence of IFN-γ ( a ), IL-4 ( b ), IL-6 ( c ) and IL-21 ( d ) by ELISA. Graphs show cytokine responses and values are expressed as mean ± SEM from each mouse from three independent infection experiments ( n = 10 IL-17A −/− and n = 13 WT mice). Statistical significances between the indicated groups were obtained using the Mann-Whitney U tests. Asterisks denote significant differences between the groups indicated by the brackets (* p

    Article Snippet: Filarial-specific cell culture assays For bulk cell assays, 5 × 105 erythrocyte-depleted mLN cells from individual mice were plated in 96-well round-bottomed plates in RPMI 1640 medium containing 10% FCS, 1% Pen/Strep, 1% L-glutamine and 0.1% gentamycin (Thermo Fisher Scientific).

    Techniques: Infection, Mouse Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY