il 4  (BioLegend)

 
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    Name:
    PE Cy7 anti human IL 4
    Description:
    PE Cy7 anti human IL 4 MP4 25D2 Isotype Rat IgG1 κ Reactivity Human Cross Reactivity Swine Pig Porcine Rhesus Apps ICFC Size 25 tests
    Catalog Number:
    500823
    Price:
    110
    Category:
    Human Cytokine Chemokine
    Source:
    Rat
    Applications:
    ICFC
    Conjugate:
    PE Cy7
    Immunogen:
    CHO-expressed, recombinant human IL-4
    Size:
    25 tests
    Quantity:
    1
    Preparation:
    The antibody was purified by affinity chromatography and conjugated with PE Cy7 under optimal conditions The solution is free of unconjugated PE Cy7 and unconjugated antibody
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    Structured Review

    BioLegend il 4
    PE Cy7 anti human IL 4
    PE Cy7 anti human IL 4 MP4 25D2 Isotype Rat IgG1 κ Reactivity Human Cross Reactivity Swine Pig Porcine Rhesus Apps ICFC Size 25 tests
    https://www.bioz.com/result/il 4/product/BioLegend
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    il 4 - by Bioz Stars, 2021-05
    94/100 stars

    Images

    1) Product Images from "Inherited salt-losing tubulopathies are associated with immunodeficiency due to impaired IL-17 responses"

    Article Title: Inherited salt-losing tubulopathies are associated with immunodeficiency due to impaired IL-17 responses

    Journal: Nature Communications

    doi: 10.1038/s41467-020-18184-3

    Effect of altering extracellular ion concentration within a physiological range on CD4 subset balance. a Effect of NaCl (0–40 mM), KCl (0–2 mM), and MgCl 2 (0–0.8 mM) during T-cell activation with anti-CD3/anti-CD28 on the ratio of expression of IFNγ:IL-17 in CD4 + cells, representative of Th1:Th17 cells. Data shown represent dose–response in a healthy control. Mean of technical duplicates is plotted. b Effect of NaCl (0–40 mM), KCl (0–2 mM), and MgCl 2 (0–0.8 mM) during T-cell activation with anti-CD3/anti-CD28 on the ratio of expression of IL-4:IL-17 in CD4 + cells, representative of Th2:Th17 cells. Data shown represent dose–response in a healthy control. Mean of technical duplicates is plotted. c Representative FACS dot plots of the effect of +40 mM NaCl on the expression of IFNγ, IL-4, and IL-17 in CD4 + cells from a healthy control. Cell proportions as percentages are documented within each quadrant. d CD4 + subset balance after T-cell activation with anti-CD3/anti-CD28 in the presence of NaCl (40 mM), KCl (2 mM), and MgCl 2 (1 mM) used alone and in combination. Data show subset balance reported as a ratio to standard conditions in healthy controls ( n = 2). The median ratio is plotted. Lines drawn at a ratio of 1 represents no difference in subset balance in the presence of ions compared to standard conditions. Source data are provided as a Source data file.
    Figure Legend Snippet: Effect of altering extracellular ion concentration within a physiological range on CD4 subset balance. a Effect of NaCl (0–40 mM), KCl (0–2 mM), and MgCl 2 (0–0.8 mM) during T-cell activation with anti-CD3/anti-CD28 on the ratio of expression of IFNγ:IL-17 in CD4 + cells, representative of Th1:Th17 cells. Data shown represent dose–response in a healthy control. Mean of technical duplicates is plotted. b Effect of NaCl (0–40 mM), KCl (0–2 mM), and MgCl 2 (0–0.8 mM) during T-cell activation with anti-CD3/anti-CD28 on the ratio of expression of IL-4:IL-17 in CD4 + cells, representative of Th2:Th17 cells. Data shown represent dose–response in a healthy control. Mean of technical duplicates is plotted. c Representative FACS dot plots of the effect of +40 mM NaCl on the expression of IFNγ, IL-4, and IL-17 in CD4 + cells from a healthy control. Cell proportions as percentages are documented within each quadrant. d CD4 + subset balance after T-cell activation with anti-CD3/anti-CD28 in the presence of NaCl (40 mM), KCl (2 mM), and MgCl 2 (1 mM) used alone and in combination. Data show subset balance reported as a ratio to standard conditions in healthy controls ( n = 2). The median ratio is plotted. Lines drawn at a ratio of 1 represents no difference in subset balance in the presence of ions compared to standard conditions. Source data are provided as a Source data file.

    Techniques Used: Concentration Assay, Activation Assay, Expressing, FACS

    CD4 subset analysis in salt-losing tubulopathy patients and controls. a IFNγ, IL-4, and IL-17 expression in CD4 + cells after 4 h stimulation with phorbol myristate acetate and ionomycin in SLT patients ( n = 15), healthy controls ( n = 11), and disease controls ( n = 12). Cytokine expression is reported as percentage of CD4 + cells. Groups are compared with a Kruskal–Wallis test with Dunn’s multiple comparison testing (shown with significance bars). Error bars represent interquartile range around the median. IFNγ expression: HC 1.9% (1.5–4.0), SLT 1.3% (1.0–3.7), DC 0.9% (0.5–8.9), p = 0.53. IL-4 expression: HC 2.8% (0.8–4.8), SLT 3.0% (1.7–3.3), DC 2.5% (0.7–3.0), p = 0.52. IL-17 expression: HC 1.4% (0.9–2.2), SLT 0.7% (0.3–1.4), DC 1.0% (0.8–1.4), p = 0.038. b Representative FACS dot plots of IFNγ, IL-4, and IL-17 expression in CD4 + cells in a SLT patient and HC. Cell proportions as percentages are documented within each quadrant. c Ratio of IFNγ, IL-4, and IL-17 expression in each subject in SLT patients ( n = 15), HC ( n = 11), and DC ( n = 12). Groups are compared with a Kruskal–Wallis test with Dunn’s multiple comparison testing (shown with significance bars). Error bars represent interquartile range around the median. IFNγ:IL-4 (Th1:Th2): HC 1.2 (0.4–2.9), SLT 0.7 (0.3–3.9), DC (1.0 (0.4–3.9), p = 0.82). IFNγ:IL-17 (Th1:Th17): HC 1.8 (0.8–7.0), SLT 3.3 (1.6–8.3), DC 1.0 (0.5–7.8), p = 0.40. IL-4:IL-17 (Th2:Th17): HC 1.9 (0.4–3.2), SLT 3.4 (1.2–6.6), DC 1.4 (0.7–2.5), p = 0.05. ns not significant ( p > 0.05), * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001, HC healthy control, SLT salt-losing tubulopathy, DC disease control. Source data are provided as a Source data file.
    Figure Legend Snippet: CD4 subset analysis in salt-losing tubulopathy patients and controls. a IFNγ, IL-4, and IL-17 expression in CD4 + cells after 4 h stimulation with phorbol myristate acetate and ionomycin in SLT patients ( n = 15), healthy controls ( n = 11), and disease controls ( n = 12). Cytokine expression is reported as percentage of CD4 + cells. Groups are compared with a Kruskal–Wallis test with Dunn’s multiple comparison testing (shown with significance bars). Error bars represent interquartile range around the median. IFNγ expression: HC 1.9% (1.5–4.0), SLT 1.3% (1.0–3.7), DC 0.9% (0.5–8.9), p = 0.53. IL-4 expression: HC 2.8% (0.8–4.8), SLT 3.0% (1.7–3.3), DC 2.5% (0.7–3.0), p = 0.52. IL-17 expression: HC 1.4% (0.9–2.2), SLT 0.7% (0.3–1.4), DC 1.0% (0.8–1.4), p = 0.038. b Representative FACS dot plots of IFNγ, IL-4, and IL-17 expression in CD4 + cells in a SLT patient and HC. Cell proportions as percentages are documented within each quadrant. c Ratio of IFNγ, IL-4, and IL-17 expression in each subject in SLT patients ( n = 15), HC ( n = 11), and DC ( n = 12). Groups are compared with a Kruskal–Wallis test with Dunn’s multiple comparison testing (shown with significance bars). Error bars represent interquartile range around the median. IFNγ:IL-4 (Th1:Th2): HC 1.2 (0.4–2.9), SLT 0.7 (0.3–3.9), DC (1.0 (0.4–3.9), p = 0.82). IFNγ:IL-17 (Th1:Th17): HC 1.8 (0.8–7.0), SLT 3.3 (1.6–8.3), DC 1.0 (0.5–7.8), p = 0.40. IL-4:IL-17 (Th2:Th17): HC 1.9 (0.4–3.2), SLT 3.4 (1.2–6.6), DC 1.4 (0.7–2.5), p = 0.05. ns not significant ( p > 0.05), * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001, HC healthy control, SLT salt-losing tubulopathy, DC disease control. Source data are provided as a Source data file.

    Techniques Used: Expressing, FACS

    Related Articles

    Flow Cytometry:

    Article Title: Lipopolysaccharide inhalation recruits monocytes and dendritic cell subsets to the alveolar airspace
    Article Snippet: Dead cells were stained with a Zombie Aqua (Biolegend) prior to fixation and permeabilization (BD) according to manufacturers instructions. .. Cells were prepared for flow cytometry as above using anti-IFN-γ-PE/Dazzle-594 (4s.B3; Biolegend; 505845; 1:100), anti-IL17-AF647 (BL168; Biolegend; 512309; 1:100) and anti-IL4-PECy7 (MP4–25D2; Biolegend; 500823; 1:100) and analyzed on a Fortessa X20 (BD) running FACSDIVA version 7. .. Ex-vivo macrophage stimulation Alveolar macrophages and CD14++ CD16− monocyte/macrophages were sorted from LPS BAL (n = 4 each).

    Article Title: Broad phenotypic alterations and potential dysfunctions of lymphocytes in COVID-19 recovered individuals
    Article Snippet: .. .https://doi.org/10.1101/2020.07.01.20144030doi: medRxiv preprint Supplementary Table 2 (Table S2): reagents for flow cytometry antibodies (clone) staining procedure source identifier Fixable Viability Dye eFluor™ 506 surface eBioscience 65-0866-14 Alexa Fluor® 700 anti-human CD183 (CXCR3) (G025H7) surface BioLegend 353742 Alexa Fluor® 700 anti-human CD3 (SK7) surface BioLegend 344822 Alexa Fluor® 700 anti-human CD4 (OKT4) surface BioLegend 317426 APC anti-human CD196 (CCR6) (G034E3) surface BioLegend 353416 APC anti-human CD275 (B7-H2, ICOSL) (2D3) surface BioLegend 309408 APC/Cyanine7 anti-human CD3 (HIT3a) surface BioLegend 300318 APC/Cyanine7 anti-human CD45RA (HI100) surface BioLegend 304127 Brilliant Violet 421™ anti-human/mouse/rat CD278 (ICOS)(C398.4A) surface BioLegend 313524 Brilliant Violet 605™ anti-human CD19 (HIB19) surface BioLegend 302244 Brilliant Violet 605™ anti-human CD8a (RPA-T8) surface BioLegend 301040 Brilliant Violet 650™ anti-human CD38 (HB-7) surface BioLegend 356620 Brilliant Violet 650™ anti-human CD4 (RPA-T4) surface BioLegend 300536 FITC anti-human CD20 (2H7) surface BioLegend 302304 FITC anti-human CD25 (BC96) surface BioLegend 302604 FITC anti-human CD45RO (UCHL1) surface BioLegend 304242 Pacific Blue™ anti-human CD27 ( O323) surface BioLegend 302822 PE anti-human IgM (MHM-88) surface BioLegend 314508 PE/Cy7 anti-human CTLA-4 (L3D10) surface BioLegend 349914 PE/Cyanine7 anti-human CD27 (O323) surface BioLegend 302838 PE/Dazzle™ 594 anti-human CD279 (PD-1 (EH12.2H7) surface BioLegend 329940 PE/Dazzle™ 594 anti-human CD71 (CY1G4) surface BioLegend 334120 PerCP/Cy5.5 anti-human CD127 (A019D5) surface BioLegend 351322 PerCP/Cyanine5.5 anti-human CD185 (CXCR5) (J252D4) surface BioLegend 356910 PerCP/Cyanine5.5 anti-human HLA-DR (L243) surface BioLegend 307630 Brilliant Violet 421™ anti-human FoxP3 (206D) intracellular BioLegend 320124 Brilliant Violet 650™ anti-human IL-2 (MQ1-17H12) intracellular BioLegend 500334 FITC anti-human/mouse Granzyme B (QA16A02) intracellular BioLegend 372206 PE anti-human IL-21 (3A3-N2) intracellular BioLegend 513004 PE anti-human Ki-67 (Ki-67) intracellular BioLegend 350504 PE/Cy7 anti-human IL-4 (MP4-25D2 ) intracellular BioLegend 500824 PE/Cyanine7 anti-human Ki-67 ( Ki-67) intracellular BioLegend 350526 PE/Dazzle™ 594 anti-human IFN-γ (B27) intracellular BioLegend 506530 PerCP/Cy5.5 anti-human IL-17A (BL168) intracellular BioLegend 512314 All rights reserved. ..

    Cytometry:

    Article Title: Lipopolysaccharide inhalation recruits monocytes and dendritic cell subsets to the alveolar airspace
    Article Snippet: Dead cells were stained with a Zombie Aqua (Biolegend) prior to fixation and permeabilization (BD) according to manufacturers instructions. .. Cells were prepared for flow cytometry as above using anti-IFN-γ-PE/Dazzle-594 (4s.B3; Biolegend; 505845; 1:100), anti-IL17-AF647 (BL168; Biolegend; 512309; 1:100) and anti-IL4-PECy7 (MP4–25D2; Biolegend; 500823; 1:100) and analyzed on a Fortessa X20 (BD) running FACSDIVA version 7. .. Ex-vivo macrophage stimulation Alveolar macrophages and CD14++ CD16− monocyte/macrophages were sorted from LPS BAL (n = 4 each).

    Staining:

    Article Title: Inherited salt-losing tubulopathies are associated with immunodeficiency due to impaired IL-17 responses
    Article Snippet: T-cell subset analysis PBMCs were isolated from SLT patients (n = 20), HCs (n = 19), and disease controls (n = 18) and fresh cells were stimulated for 4 h with phorbol myristate acetate (PMA, 50 ng/ml; Sigma, cat#P8139) and ionomycin (1 µg/ml; Sigma, cat#I9657) in the presence of brefeldin A (5 µg/ml). .. Cells were subsequently stained for CD4 (CD4-FITC), IL-17 (IL-17-PE; BioLegend, cat#512306), IFNγ (IFNγ-APC; BioLegend, cat#502512) and IL-4 (IL-4-PE/Cy7; BioLegend; cat500824), and analysed by FACS. .. The proportion of CD4+ cells expressing each of these cytokines was determined and compared between groups.

    Article Title: The Protective Effects of IL-31RA Deficiency During Bleomycin-Induced Pulmonary Fibrosis
    Article Snippet: Non-specific binding of fluorochromes was prevented by incubating cells with Human BD Fc Block™ (BD Biosciences), followed by surface marker staining with anti-CD3-V500 (UCHT1) and anti-CD4-APC(RPA-T4) (Bio Legend). .. Cells were then fixed and permeabilized using Cytofix/Cytoperm (BD Biosciences) and stained with antibodies against IL-31- PE (U26-947) (BD Biosciences) and IL-4 -PE-Cy7 (MP4-25D2) (BioLegend). .. Data was acquired using LSR Fortessa flow cytometer and analyzed using FlowJo 10 (FlowJo, Tree Star).

    Article Title: Broad phenotypic alterations and potential dysfunctions of lymphocytes in COVID-19 recovered individuals
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    Article Title: Inherited salt-losing tubulopathies are associated with immunodeficiency due to impaired IL-17 responses
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    FACS:

    Article Title: Inherited salt-losing tubulopathies are associated with immunodeficiency due to impaired IL-17 responses
    Article Snippet: T-cell subset analysis PBMCs were isolated from SLT patients (n = 20), HCs (n = 19), and disease controls (n = 18) and fresh cells were stimulated for 4 h with phorbol myristate acetate (PMA, 50 ng/ml; Sigma, cat#P8139) and ionomycin (1 µg/ml; Sigma, cat#I9657) in the presence of brefeldin A (5 µg/ml). .. Cells were subsequently stained for CD4 (CD4-FITC), IL-17 (IL-17-PE; BioLegend, cat#512306), IFNγ (IFNγ-APC; BioLegend, cat#502512) and IL-4 (IL-4-PE/Cy7; BioLegend; cat500824), and analysed by FACS. .. The proportion of CD4+ cells expressing each of these cytokines was determined and compared between groups.

    Article Title: Inherited salt-losing tubulopathies are associated with immunodeficiency due to impaired IL-17 responses
    Article Snippet: On day 3, cells were restimulated for 4 h with PMA (50 ng/ml) and ionomycin (1 µg/ml; Sigma, cat#I9657) in the presence of brefeldin A (5 µg/ml). .. Cells were then stained for viability (eFluor450), CD4 (CD4-FITC), IFNγ (IFNγ-APC), IL-4 (IL-4-PE/Cy7) and IL-17 (IL-17-PE), and analysed by FACS. .. The ratio of cytokine expression in viable CD4+ cells was determined.

    Recombinase Polymerase Amplification:

    Article Title: Broad phenotypic alterations and potential dysfunctions of lymphocytes in COVID-19 recovered individuals
    Article Snippet: .. .https://doi.org/10.1101/2020.07.01.20144030doi: medRxiv preprint Supplementary Table 2 (Table S2): reagents for flow cytometry antibodies (clone) staining procedure source identifier Fixable Viability Dye eFluor™ 506 surface eBioscience 65-0866-14 Alexa Fluor® 700 anti-human CD183 (CXCR3) (G025H7) surface BioLegend 353742 Alexa Fluor® 700 anti-human CD3 (SK7) surface BioLegend 344822 Alexa Fluor® 700 anti-human CD4 (OKT4) surface BioLegend 317426 APC anti-human CD196 (CCR6) (G034E3) surface BioLegend 353416 APC anti-human CD275 (B7-H2, ICOSL) (2D3) surface BioLegend 309408 APC/Cyanine7 anti-human CD3 (HIT3a) surface BioLegend 300318 APC/Cyanine7 anti-human CD45RA (HI100) surface BioLegend 304127 Brilliant Violet 421™ anti-human/mouse/rat CD278 (ICOS)(C398.4A) surface BioLegend 313524 Brilliant Violet 605™ anti-human CD19 (HIB19) surface BioLegend 302244 Brilliant Violet 605™ anti-human CD8a (RPA-T8) surface BioLegend 301040 Brilliant Violet 650™ anti-human CD38 (HB-7) surface BioLegend 356620 Brilliant Violet 650™ anti-human CD4 (RPA-T4) surface BioLegend 300536 FITC anti-human CD20 (2H7) surface BioLegend 302304 FITC anti-human CD25 (BC96) surface BioLegend 302604 FITC anti-human CD45RO (UCHL1) surface BioLegend 304242 Pacific Blue™ anti-human CD27 ( O323) surface BioLegend 302822 PE anti-human IgM (MHM-88) surface BioLegend 314508 PE/Cy7 anti-human CTLA-4 (L3D10) surface BioLegend 349914 PE/Cyanine7 anti-human CD27 (O323) surface BioLegend 302838 PE/Dazzle™ 594 anti-human CD279 (PD-1 (EH12.2H7) surface BioLegend 329940 PE/Dazzle™ 594 anti-human CD71 (CY1G4) surface BioLegend 334120 PerCP/Cy5.5 anti-human CD127 (A019D5) surface BioLegend 351322 PerCP/Cyanine5.5 anti-human CD185 (CXCR5) (J252D4) surface BioLegend 356910 PerCP/Cyanine5.5 anti-human HLA-DR (L243) surface BioLegend 307630 Brilliant Violet 421™ anti-human FoxP3 (206D) intracellular BioLegend 320124 Brilliant Violet 650™ anti-human IL-2 (MQ1-17H12) intracellular BioLegend 500334 FITC anti-human/mouse Granzyme B (QA16A02) intracellular BioLegend 372206 PE anti-human IL-21 (3A3-N2) intracellular BioLegend 513004 PE anti-human Ki-67 (Ki-67) intracellular BioLegend 350504 PE/Cy7 anti-human IL-4 (MP4-25D2 ) intracellular BioLegend 500824 PE/Cyanine7 anti-human Ki-67 ( Ki-67) intracellular BioLegend 350526 PE/Dazzle™ 594 anti-human IFN-γ (B27) intracellular BioLegend 506530 PerCP/Cy5.5 anti-human IL-17A (BL168) intracellular BioLegend 512314 All rights reserved. ..

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    BioLegend anti mouse il 4
    IL-10 replicates IL-6 mediated Th1/Th2 skewing and IL-21 expression in IL-10Rα expressing cells (A) CD4 + cells from spleens and lymph nodes of IL-10Rα and vector retrogenic mice were sorted based on GFP expression and cultured under the indicated conditions for 5 days, restimulated with PMA/ionomycin/brefeldin A, and stained for intracellular IFN-γ (top) and <t>IL-4</t> (bottom). Graphs are representative of 3 or more independent experiments and normalized to the maximal percent of cytokine-expressing cells. (B) CD4 + cells from spleen and lymph nodes of IL-10Rα retrogenic mice were sorted based on GFP expression and cultured in the presence of irradiated APCs and anti-CD3 antibody with the indicated cytokines for 4 days. Media containing added cytokines was removed by washing and replaced with fresh media for 24 hours. Supernatants were assayed for IL-21 by ELISA. ** p≤ 0.01; *** p≤ 0.005; **** p≤ 0.0001.
    Anti Mouse Il 4, supplied by BioLegend, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    P4 is an effective suppressor of T cell differentiation into Th17 cells Treg-depleted CB T cells were cultured in a Th17 cell-induction condition (anti-CD3/CD28 beads or PHA in the presence of IL-23, IL-1β, TGFβ1, <t>anti-IL-4,</t> and anti-IFNγ) for 7 days with or without P4 (2 µg/ml). The relative frequencies of Th17 cells and Th1 cells (A) and expression of Th17-associated genes at the RNA level determined with a real-time PCR method (B) is shown. The combined data in panel A were obtained from 8–12, and the data in panel B from 3–5, independent experiments. *Significant differences from the control group.
    Anti Il 4, supplied by BioLegend, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Calpastatin-overexpression in Th cells resulted in decreased IL-6 and IL-17 production. (A) Naïve CD4 + T cells were cultured under neutral, Th1, Th2, or Th17 conditions. On day 1, these cells were infected with mock-GRV or calpain- or CS-dIV-GRV and expanded on day 2 with IL-2. On day 4 (Th17 conditions) or on day 6 (neutral, Th1, and Th2 conditions), the cells were restimulated with PMA/ionomycin for ICS. Representative results for GFP + -gated cells are shown (n = 11 for IFNγ, n = 10 for <t>IL-4,</t> n = 6 for IL-6 and IL-17). (B) Statistical analysis of the results in panel A by Steel's test. (C) Naïve CD4 + T cells were cultured under neutral conditions and infected with mock-GRV or CS-dIV-GRV. On day 6, these cells were restimulated with PMA/ionomycin for FACS. Representative data of GFP + , GFP − -gated cells are shown in the left panel, and the statistical analysis is shown in the right panel (n = 3) (D) Retrovirus-infected Th cells were stained with PKH-26 dye and then stimulated with anti-CD3/28 monoclonal antibodies. The fluorescence intensity of the PKH-26 dye in infected (GFP + ) and non-infected (GFP − ) cells was analyzed by FACS. *p
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    IL-10 replicates IL-6 mediated Th1/Th2 skewing and IL-21 expression in IL-10Rα expressing cells (A) CD4 + cells from spleens and lymph nodes of IL-10Rα and vector retrogenic mice were sorted based on GFP expression and cultured under the indicated conditions for 5 days, restimulated with PMA/ionomycin/brefeldin A, and stained for intracellular IFN-γ (top) and IL-4 (bottom). Graphs are representative of 3 or more independent experiments and normalized to the maximal percent of cytokine-expressing cells. (B) CD4 + cells from spleen and lymph nodes of IL-10Rα retrogenic mice were sorted based on GFP expression and cultured in the presence of irradiated APCs and anti-CD3 antibody with the indicated cytokines for 4 days. Media containing added cytokines was removed by washing and replaced with fresh media for 24 hours. Supernatants were assayed for IL-21 by ELISA. ** p≤ 0.01; *** p≤ 0.005; **** p≤ 0.0001.

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: Differential T cell cytokine receptivity and not signal quality distinguishes IL-6 and IL-10 signaling during Th17 differentiation

    doi: 10.4049/jimmunol.1402953

    Figure Lengend Snippet: IL-10 replicates IL-6 mediated Th1/Th2 skewing and IL-21 expression in IL-10Rα expressing cells (A) CD4 + cells from spleens and lymph nodes of IL-10Rα and vector retrogenic mice were sorted based on GFP expression and cultured under the indicated conditions for 5 days, restimulated with PMA/ionomycin/brefeldin A, and stained for intracellular IFN-γ (top) and IL-4 (bottom). Graphs are representative of 3 or more independent experiments and normalized to the maximal percent of cytokine-expressing cells. (B) CD4 + cells from spleen and lymph nodes of IL-10Rα retrogenic mice were sorted based on GFP expression and cultured in the presence of irradiated APCs and anti-CD3 antibody with the indicated cytokines for 4 days. Media containing added cytokines was removed by washing and replaced with fresh media for 24 hours. Supernatants were assayed for IL-21 by ELISA. ** p≤ 0.01; *** p≤ 0.005; **** p≤ 0.0001.

    Article Snippet: IL-23R and CCR6Low endotoxin, azide-free neutralizing anti-mouse IL-4 (clone 11B11) and anti-mouse IFN-γ (XMG1.2) were obtained from Biolegend.

    Techniques: Expressing, Plasmid Preparation, Mouse Assay, Cell Culture, Staining, Irradiation, Enzyme-linked Immunosorbent Assay

    P4 is an effective suppressor of T cell differentiation into Th17 cells Treg-depleted CB T cells were cultured in a Th17 cell-induction condition (anti-CD3/CD28 beads or PHA in the presence of IL-23, IL-1β, TGFβ1, anti-IL-4, and anti-IFNγ) for 7 days with or without P4 (2 µg/ml). The relative frequencies of Th17 cells and Th1 cells (A) and expression of Th17-associated genes at the RNA level determined with a real-time PCR method (B) is shown. The combined data in panel A were obtained from 8–12, and the data in panel B from 3–5, independent experiments. *Significant differences from the control group.

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: Progesterone promotes differentiation of human cord blood fetal T cells into T regulatory cells but suppresses their differentiation into Th17 cells 1

    doi: 10.4049/jimmunol.1003919

    Figure Lengend Snippet: P4 is an effective suppressor of T cell differentiation into Th17 cells Treg-depleted CB T cells were cultured in a Th17 cell-induction condition (anti-CD3/CD28 beads or PHA in the presence of IL-23, IL-1β, TGFβ1, anti-IL-4, and anti-IFNγ) for 7 days with or without P4 (2 µg/ml). The relative frequencies of Th17 cells and Th1 cells (A) and expression of Th17-associated genes at the RNA level determined with a real-time PCR method (B) is shown. The combined data in panel A were obtained from 8–12, and the data in panel B from 3–5, independent experiments. *Significant differences from the control group.

    Article Snippet: For induction of Th17 cells, CB naive CD4+ T cells were activated with anti-CD3/CD28 beads (Miltenyi Biotec Inc) for 6–7 days in RPMI medium supplemented with FBS (10%), IL-23 (25 ng/ml; R & D Systems), IL-1β (10 ng/ml; PeproTech, Inc. Rocky Hill, NJ), TGFβ1 (1 ng/ml; PeproTech), anti-IL-4 (10 µg/ml; MP4-25D2, BioLegend Inc), and anti-IFNγ (10 µg/ml; MD-1, BioLegend Inc).

    Techniques: Cell Differentiation, Cell Culture, Expressing, Real-time Polymerase Chain Reaction

    Calpastatin-overexpression in Th cells resulted in decreased IL-6 and IL-17 production. (A) Naïve CD4 + T cells were cultured under neutral, Th1, Th2, or Th17 conditions. On day 1, these cells were infected with mock-GRV or calpain- or CS-dIV-GRV and expanded on day 2 with IL-2. On day 4 (Th17 conditions) or on day 6 (neutral, Th1, and Th2 conditions), the cells were restimulated with PMA/ionomycin for ICS. Representative results for GFP + -gated cells are shown (n = 11 for IFNγ, n = 10 for IL-4, n = 6 for IL-6 and IL-17). (B) Statistical analysis of the results in panel A by Steel's test. (C) Naïve CD4 + T cells were cultured under neutral conditions and infected with mock-GRV or CS-dIV-GRV. On day 6, these cells were restimulated with PMA/ionomycin for FACS. Representative data of GFP + , GFP − -gated cells are shown in the left panel, and the statistical analysis is shown in the right panel (n = 3) (D) Retrovirus-infected Th cells were stained with PKH-26 dye and then stimulated with anti-CD3/28 monoclonal antibodies. The fluorescence intensity of the PKH-26 dye in infected (GFP + ) and non-infected (GFP − ) cells was analyzed by FACS. *p

    Journal: PLoS ONE

    Article Title: Overexpression of a Minimal Domain of Calpastatin Suppresses IL-6 Production and Th17 Development via Reduced NF-?B and Increased STAT5 Signals

    doi: 10.1371/journal.pone.0027020

    Figure Lengend Snippet: Calpastatin-overexpression in Th cells resulted in decreased IL-6 and IL-17 production. (A) Naïve CD4 + T cells were cultured under neutral, Th1, Th2, or Th17 conditions. On day 1, these cells were infected with mock-GRV or calpain- or CS-dIV-GRV and expanded on day 2 with IL-2. On day 4 (Th17 conditions) or on day 6 (neutral, Th1, and Th2 conditions), the cells were restimulated with PMA/ionomycin for ICS. Representative results for GFP + -gated cells are shown (n = 11 for IFNγ, n = 10 for IL-4, n = 6 for IL-6 and IL-17). (B) Statistical analysis of the results in panel A by Steel's test. (C) Naïve CD4 + T cells were cultured under neutral conditions and infected with mock-GRV or CS-dIV-GRV. On day 6, these cells were restimulated with PMA/ionomycin for FACS. Representative data of GFP + , GFP − -gated cells are shown in the left panel, and the statistical analysis is shown in the right panel (n = 3) (D) Retrovirus-infected Th cells were stained with PKH-26 dye and then stimulated with anti-CD3/28 monoclonal antibodies. The fluorescence intensity of the PKH-26 dye in infected (GFP + ) and non-infected (GFP − ) cells was analyzed by FACS. *p

    Article Snippet: Anti-IL-4 antibody (11B11) was purchased from BioLegend Inc., and anti-IFN-γ antibody (XMG1.2) from BD Biosciences.

    Techniques: Over Expression, Cell Culture, Infection, FACS, Staining, Fluorescence

    Effect of CBD on IL-2 and TGF-β1 production Splenocytes were treated with CBD (5 or 10 μM) or VH (0.1% ethanol) for 30 min followed by low-level stimulation. (A) Cells were stimulated with S/o or Us/o stimulation for 1, 3 or 5 days. IL-2 was assessed by ELISA. Data are average of triplicate wells ± SD with * p

    Journal: Cellular immunology

    Article Title: Cannabidiol (CBD) Induces Functional Tregs in Response to Low-Level T Cell Activation

    doi: 10.1016/j.cellimm.2016.11.006

    Figure Lengend Snippet: Effect of CBD on IL-2 and TGF-β1 production Splenocytes were treated with CBD (5 or 10 μM) or VH (0.1% ethanol) for 30 min followed by low-level stimulation. (A) Cells were stimulated with S/o or Us/o stimulation for 1, 3 or 5 days. IL-2 was assessed by ELISA. Data are average of triplicate wells ± SD with * p

    Article Snippet: Wells were coated overnight at 4°C with 1.0 μg/ml anti-mouse IL-2 (clone JES6-1A12, BioLegend) in coating buffer (0.1 M carbonate-bicarbonate buffer, pH 9.6).

    Techniques: Enzyme-linked Immunosorbent Assay