elisa kits  (Thermo Fisher)


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    Name:
    IL 6 Mouse ELISA Kit
    Description:
    This Mouse IL 6 Platinum ELISA contains the necessary reagents from pre coated plates to standards buffers and diluents for performing quantitative enzyme linked immunosorbent assays ELISA This ELISA kit is specifically engineered for accurate and precise measurement of IL 6 protein levels from samples including serum plasma and supernatants from cell cultures Interleukin 6 IL 6 is a multi functional cytokine that regulates immune responses acute phase reactions and hematopoiesis and may play a central role in host defense mechanisms IL 6 is usually not produced constitutively by normal cells but its expression is readily induced by a variety of cytokines lipopolysaccharide LPS or viral infections IL 6 is a pleiotropic cytokine produced by a variety of cells It acts on a wide range of tissues exerting growth induction growth inhibition and differentiation respectively depending on the nature of the target cells IL 6 is involved in the induction of B cell differentiation the induction of acute phase proteins in liver cells growth promotion of myeloma plasmacytoma hybridoma cells induction of IL 2 and IL 2 receptor expression proliferation and differentiation of T cells inhibition of cell growth of certain myeloid leukemic cell lines and induction of their differentiation to macrophages enhancement of IL 3 induced multipotential colony cell formation in hematopoietic stem cells and induction of maturation of megakaryocytes as a thrombopoietic factor induction of mesangial cell growth induction of neural differentiation of PC 12 cells induction of keratinocyte growth The abnormal production of IL 6 was first suggested to be related to polyclonal B cell activation with autoantibody production in patients with cardiac myxoma Since then IL 6 has been suggested to be involved in the pathogenesis of a variety of diseases Measurement of IL 6 levels in serum and other body fluids thus provides more detailed insights into various pathological situations ConjugateBiotinSuitable Sample Typescell culture supernatant serum plasma EDTA citrate Sample Volume50 µLReported ApplicationELISA
    Catalog Number:
    BMS603-2
    Price:
    None
    Category:
    Kits and Assays
    Applications:
    Protein Assays and Analysis|Protein Biology
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    Structured Review

    Thermo Fisher elisa kits
    The inflammatory response of HBeAg‐induced macrophages was inhibited by miR‐212‐3p. miR‐212‐3p mimics and negative control were transfected into RAW264.7 macrophages for 24 h, then the cells were treated with HBeAg for 24 h, and the expression of miR‐212‐3p (A) and level of IL‐6 (B, C), <t>TNF‐α</t> (D, E) were tested by q‐PCR and <t>ELISA.</t> miR‐212‐3p inhibitor and negative control were transfected into RAW264.7 macrophages for 24 h, then the cells were treated with HBeAg for 24 h; and the expression of miR‐212‐3p (F) and level of IL‐6 (G, H), TNF‐α (I, J) were tested by q‐PCR and ELISA. Data are shown in at least three independent experiments (mean ± SD of triplicates in A‐J). * P
    This Mouse IL 6 Platinum ELISA contains the necessary reagents from pre coated plates to standards buffers and diluents for performing quantitative enzyme linked immunosorbent assays ELISA This ELISA kit is specifically engineered for accurate and precise measurement of IL 6 protein levels from samples including serum plasma and supernatants from cell cultures Interleukin 6 IL 6 is a multi functional cytokine that regulates immune responses acute phase reactions and hematopoiesis and may play a central role in host defense mechanisms IL 6 is usually not produced constitutively by normal cells but its expression is readily induced by a variety of cytokines lipopolysaccharide LPS or viral infections IL 6 is a pleiotropic cytokine produced by a variety of cells It acts on a wide range of tissues exerting growth induction growth inhibition and differentiation respectively depending on the nature of the target cells IL 6 is involved in the induction of B cell differentiation the induction of acute phase proteins in liver cells growth promotion of myeloma plasmacytoma hybridoma cells induction of IL 2 and IL 2 receptor expression proliferation and differentiation of T cells inhibition of cell growth of certain myeloid leukemic cell lines and induction of their differentiation to macrophages enhancement of IL 3 induced multipotential colony cell formation in hematopoietic stem cells and induction of maturation of megakaryocytes as a thrombopoietic factor induction of mesangial cell growth induction of neural differentiation of PC 12 cells induction of keratinocyte growth The abnormal production of IL 6 was first suggested to be related to polyclonal B cell activation with autoantibody production in patients with cardiac myxoma Since then IL 6 has been suggested to be involved in the pathogenesis of a variety of diseases Measurement of IL 6 levels in serum and other body fluids thus provides more detailed insights into various pathological situations ConjugateBiotinSuitable Sample Typescell culture supernatant serum plasma EDTA citrate Sample Volume50 µLReported ApplicationELISA
    https://www.bioz.com/result/elisa kits/product/Thermo Fisher
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    elisa kits - by Bioz Stars, 2021-05
    95/100 stars

    Images

    1) Product Images from "Negative feedback loop of ERK/CREB/miR‐212‐3p inhibits HBeAg‐induced macrophage activation, et al. Negative feedback loop of ERK/CREB/miR‐212‐3p inhibits HBeAg‐induced macrophage activation"

    Article Title: Negative feedback loop of ERK/CREB/miR‐212‐3p inhibits HBeAg‐induced macrophage activation, et al. Negative feedback loop of ERK/CREB/miR‐212‐3p inhibits HBeAg‐induced macrophage activation

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/jcmm.15723

    The inflammatory response of HBeAg‐induced macrophages was inhibited by miR‐212‐3p. miR‐212‐3p mimics and negative control were transfected into RAW264.7 macrophages for 24 h, then the cells were treated with HBeAg for 24 h, and the expression of miR‐212‐3p (A) and level of IL‐6 (B, C), TNF‐α (D, E) were tested by q‐PCR and ELISA. miR‐212‐3p inhibitor and negative control were transfected into RAW264.7 macrophages for 24 h, then the cells were treated with HBeAg for 24 h; and the expression of miR‐212‐3p (F) and level of IL‐6 (G, H), TNF‐α (I, J) were tested by q‐PCR and ELISA. Data are shown in at least three independent experiments (mean ± SD of triplicates in A‐J). * P
    Figure Legend Snippet: The inflammatory response of HBeAg‐induced macrophages was inhibited by miR‐212‐3p. miR‐212‐3p mimics and negative control were transfected into RAW264.7 macrophages for 24 h, then the cells were treated with HBeAg for 24 h, and the expression of miR‐212‐3p (A) and level of IL‐6 (B, C), TNF‐α (D, E) were tested by q‐PCR and ELISA. miR‐212‐3p inhibitor and negative control were transfected into RAW264.7 macrophages for 24 h, then the cells were treated with HBeAg for 24 h; and the expression of miR‐212‐3p (F) and level of IL‐6 (G, H), TNF‐α (I, J) were tested by q‐PCR and ELISA. Data are shown in at least three independent experiments (mean ± SD of triplicates in A‐J). * P

    Techniques Used: Negative Control, Transfection, Expressing, Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

    MAPK pathway was activated in HBeAg‐stimulating macrophages and mediated the production of inflammatory cytokines. (A) After RAW264.7 macrophages treated with HBeAg for various times (0, 30, 60, 120, 180 min), the expression of non‐ phosphorylation and phosphorylation of ERK, JNK and p38 was detected by Western blot. RAW264.7 macrophages were pre‐treated with DMSO or inhibitor of ERK(PD98059 20 μmol/L), JNK (SP600125 20 μmol/L) and P38(SB203580 20 μmol/L) for 1 h, respectively, and then stimulated with HBeAg for 4 h, and the expression and production of IL‐6 (B, C) and TNF‐α (D, E) were examined with q‐PCR and ELISA. Data are representative of three or four independent experiments (mean ± SD of triplicates in B‐E). * P
    Figure Legend Snippet: MAPK pathway was activated in HBeAg‐stimulating macrophages and mediated the production of inflammatory cytokines. (A) After RAW264.7 macrophages treated with HBeAg for various times (0, 30, 60, 120, 180 min), the expression of non‐ phosphorylation and phosphorylation of ERK, JNK and p38 was detected by Western blot. RAW264.7 macrophages were pre‐treated with DMSO or inhibitor of ERK(PD98059 20 μmol/L), JNK (SP600125 20 μmol/L) and P38(SB203580 20 μmol/L) for 1 h, respectively, and then stimulated with HBeAg for 4 h, and the expression and production of IL‐6 (B, C) and TNF‐α (D, E) were examined with q‐PCR and ELISA. Data are representative of three or four independent experiments (mean ± SD of triplicates in B‐E). * P

    Techniques Used: Expressing, Western Blot, Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

    miR‐212‐3p regulated HBeAg‐inducing inflammatory cytokines production via targeting MAPK1. Changes in non‐ phosphorylation and phosphorylation of ERK in RAW264.7 macrophages after transfection with miR‐212‐3p mimics or miR‐212‐3p inhibitor were determined with Western blot analysis (A, B). miR‐212‐3p inhibitor or negative control miRNA were transfected into RAW264.7 for 40 h, then cells were treated with DMSO or inhibitor of ERK (PD98059, 10 μmol/L) for 2 h, and cells were stimulated with HBeAg for 4 h. The expression and production of IL‐6 (C, D) and TNF‐α (E, F) were evaluated with q‐PCR and ELISA. Columns 1 and 2 represent the change of cytokine expression and secretion with or without HBeAg stimulation after transfection of NC, respectively (C‐F). Data are shown by three or four independent experiments (mean ± SD of triplicates in C‐F). * P
    Figure Legend Snippet: miR‐212‐3p regulated HBeAg‐inducing inflammatory cytokines production via targeting MAPK1. Changes in non‐ phosphorylation and phosphorylation of ERK in RAW264.7 macrophages after transfection with miR‐212‐3p mimics or miR‐212‐3p inhibitor were determined with Western blot analysis (A, B). miR‐212‐3p inhibitor or negative control miRNA were transfected into RAW264.7 for 40 h, then cells were treated with DMSO or inhibitor of ERK (PD98059, 10 μmol/L) for 2 h, and cells were stimulated with HBeAg for 4 h. The expression and production of IL‐6 (C, D) and TNF‐α (E, F) were evaluated with q‐PCR and ELISA. Columns 1 and 2 represent the change of cytokine expression and secretion with or without HBeAg stimulation after transfection of NC, respectively (C‐F). Data are shown by three or four independent experiments (mean ± SD of triplicates in C‐F). * P

    Techniques Used: Transfection, Western Blot, Negative Control, Expressing, Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

    Related Articles

    Enzyme-linked Immunosorbent Assay:

    Article Title: Cytochrome P450 CYP2E1 Suppression Ameliorates Cerebral Ischemia Reperfusion Injury
    Article Snippet: Immunohistochemistry protocols and antibodies are presented in Materials and Methods in the Data Supplement. .. Enzyme-Linked Immunosorbent Essay Analysis (ELISA)Protein levels of inflammatory markers; tumor necrosis factor (TNF)-α, interleukin (IL)-6, and monocyte chemoattractant protein 1 (MCP-1) in the brain were measured using an ELISA kit (KMC0061c, BMS607-21NST, BMS6005 respectively, Thermo Fisher Scientific, San Jose, CA, USA) according to the manufacturer’s protocol. ..

    Article Title: Murine Coronavirus Replication-Induced p38 Mitogen-Activated Protein Kinase Activation Promotes Interleukin-6 Production and Virus Replication in Cultured Cells
    Article Snippet: .. Release of interleukin-6 (IL-6) was determined by mouse IL-6-specific enzyme-linked immunosorbent assay (ELISA) assay kit (Biosource International, Camarillo, Calif.). ..

    Article Title: Dietary Blue Pigments Derived from Genipin, Attenuate Inflammation by Inhibiting LPS-Induced iNOS and COX-2 Expression via the NF-?B Inactivation
    Article Snippet: Prostaglandin E2 Express EIA Monoclonal Kit was obtained from Cayman Chemical (USA). .. IL-6 Mouse ELISA Kit and TNF-α Mouse ELISA Kit were obtained from Invitrogen (USA). .. 4-amino-5-methylamino- 2′, 7′-difluorofluorescein diacetate (DAF-FM diacetate) was purchased from Invitrogen (USA).

    Article Title: Pathological conversion of regulatory T cells is associated with loss of allotolerance
    Article Snippet: Enzyme-linked immunosorbent assay (ELISA) The supernatant of the tissue explant culture was kept frozen at −80 °C until used for ELISA. .. ELISA detection kits for murine IL-6 and IL-23 (BMS603HS and BMS6017, eBiosciences) were used according to the manufacturer’s instructions. .. Absorbance was determined using a POLARstar Optima plate reader (BMG Labtech).

    Article Title: Negative feedback loop of ERK/CREB/miR‐212‐3p inhibits HBeAg‐induced macrophage activation, et al. Negative feedback loop of ERK/CREB/miR‐212‐3p inhibits HBeAg‐induced macrophage activation
    Article Snippet: The LightCycler Real‐time PCR System (Roche Diagnostics, Indianapolis, IN, USA) was used to perform the real‐time PCR analyses. .. 2.5 Enzyme‑linked immunosorbent assay (ELISA)We collected the cell‐culture supernatants, and the concentrations of TNF‐α (KMC3011) and IL‐6 (KMC0061) were detected using the commercially available ELISA kits (Invitrogen, Carlsbad, CA, USA). .. 2.6 Western blotAfter washed with cold phosphate‐buffered saline (PBS) for two times, the cultured cells were lysed with RIPA lysis buffer (Beyotime Biotechnology, Shanghai, China) supplemented with a cocktail of protease inhibitors.

    Article Title: Differential effect of angiotensin II and blood pressure on hippocampal inflammation in mice
    Article Snippet: .. TNF-α and IL-6 ELISA TNF-α and IL-6 were assessed by ELISA (mouse TNF-α kit #KMC3011 and mouse IL-6 kit #KMC0061, Invitrogen-Thermo Fisher Scientific, Burlington, Canada) in the brain or hippocampal homogenates as well as in plasma samples, as described in Additional file : Supplementary Methods. ..

    Article Title: The anti-inflammatory activities of ethanol extract from Dan-Lou prescription in vivo and in vitro
    Article Snippet: Dulbecco’s modified Eagle’s medium-high glucose (DMEM), 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT), aspirin and LPS from Escherichia coli 0111:B4 were obtained from Sigma-Aldrich Co. (St. Louis, USA). .. IL-6 Mouse ELISA Kit was obtained from eBioscience (San Diedo, USA). .. Mammalian Cell Lysis Kit and UNIQ-10 column Trizol total RNA extraction kit were obtained from Sangon Biological Engineering Technology & Services Co., Ltd (Shanghai, China).

    Cell Culture:

    Article Title: Negative feedback loop of ERK/CREB/miR‐212‐3p inhibits HBeAg‐induced macrophage activation, et al. Negative feedback loop of ERK/CREB/miR‐212‐3p inhibits HBeAg‐induced macrophage activation
    Article Snippet: The LightCycler Real‐time PCR System (Roche Diagnostics, Indianapolis, IN, USA) was used to perform the real‐time PCR analyses. .. 2.5 Enzyme‑linked immunosorbent assay (ELISA)We collected the cell‐culture supernatants, and the concentrations of TNF‐α (KMC3011) and IL‐6 (KMC0061) were detected using the commercially available ELISA kits (Invitrogen, Carlsbad, CA, USA). .. 2.6 Western blotAfter washed with cold phosphate‐buffered saline (PBS) for two times, the cultured cells were lysed with RIPA lysis buffer (Beyotime Biotechnology, Shanghai, China) supplemented with a cocktail of protease inhibitors.

    Article Title: Linear ubiquitin assembly complex regulates lung epithelial–driven responses during influenza infection
    Article Snippet: .. Cytokine determination.ELISAs were carried out on cell culture supernatants or BALF according to the manufacturer’s instructions using the following kits: IL-6(h) (Thermo Fisher Scientific, KHC0061C), IFN-β(h) (PBL Assay Science, 41410), IL-6(m) (Thermo Fisher Scientific, KMC0062), MCP-1(m) (Thermo Fisher Scientific, EMMCP1), IFN-α(m) (PBL Assay Science, 42120), IFN-β(m) (PBL Assay Science, 42400), IFN-γ(m) (Thermo Fisher Scientific, BMS606), and IL-10(m) (Thermo Fisher Scientific, EM2IL10). .. ChIP assay.Chromatin immunoprecipitation (ChIP) was performed using the Simple ChIP Enzymatic Chromatin IP kit (Cell Signaling, 9003) according to the manufacturer’s instructions.

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  • 93
    Thermo Fisher il 10 fitc
    Rebamipide reciprocally regulates Th17/Treg cell imbalance. Spleen tissues were isolated from mice in each group at 14 weeks after curdlan injection. a The tissues were stained with specific Abs against CD4 ( green ), CD25 ( white ), IL-17 ( red ), FOXP3 ( green ), p -STAT3–Y705 ( red ), and p -STAT3–S727 ( red ). Positive cells are shown in the bar graphs . b Spleen tissues were isolated from mice in each group at 14 weeks after curdlan injection. Splenocytes were stained with Abs against CD4–PerCP, IL-17–PE, and <t>IL-10–FITC</t> to determine the presence of Th17 and IL-10-expressing Treg cells. Data are expressed as the mean ± SEM of 3 independent experiments (* P
    Il 10 Fitc, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 93 stars, based on 1 article reviews
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    99
    Thermo Fisher cytokine
    Leishmania Ag–stimulated <t>cytokine</t> profiles in splenocytes culture supernatants from naive, LdCen-/- immunized (Imm), naive challenged (Naive Chal), and LdCen-/- immunized challenged (Imm Chal) young and aged mice. The 5-wk post immunized mice were challenged with virulent L . donovani for 4-wk and then mice were euthanized, splenocytes were isolated, plated aseptically (2×10 5 cells/well), and stimulated with Leishmania FTAg for 72h. Concentrations of pro-inflammatory cytokines IFN-γ (A), IL-12 (B) and TNF (C) anti-inflammatory cytokines IL-10 (D) and IL-4 (E) were measured in culture supernatants using the multiplex mouse cytokine kit as described in the Material and Methods section. Ratio of IFN-γ/IL-10 (F) and IFN-γ/IL-4 (G) were also determined. The data presented are representative of two independent experiments with similar results (n = 6). Mean and SEM of each group are shown. * p
    Cytokine, supplied by Thermo Fisher, 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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    99
    Thermo Fisher tnf α
    Indoleamine 2,3 dioxygenase-1 (IDO1) influences the gene expression of cytokines and transcription factors. Relative expression of mRNA for aryl hydrocarbon receptor (AhR), IFN-γ, <t>TNF-α,</t> IL-6, RORC, Tbet, GATA3, FoxP3, IL-10, TGF-β, IL-17, and IL-22 in whole lung cells of wild-type and IDO1 −/− mice after 10 weeks of Paracoccidioides brasiliensis infection. The level of gene transcription was determined by real-time PCR. Bars show mean ± SEM from at least four mice per group and are representative of three independent experiments (* p
    Tnf α, supplied by Thermo Fisher, 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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    93
    Thermo Fisher mouse il 6 elisa kit
    <t>IL-6</t> downregulation in SOD1-G93A mouse microglia. ( a ) Semiquantitative RT-PCR using specific primers for IL-6 and β -actin mRNAs, on total RNA from nt and SOD1-G93A microglia. β -actin was used for normalization. ( b ) Western blotting with anti-IL-6 antibody on total lysates from nt and SOD1-G93A microglia. β -actin was used for protein normalization. ( c ) IL-6 levels in the culture media of microglia from nt and G93A mice, after 6 h incubation in fresh media, as assessed by <t>ELISA</t>
    Mouse Il 6 Elisa Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Rebamipide reciprocally regulates Th17/Treg cell imbalance. Spleen tissues were isolated from mice in each group at 14 weeks after curdlan injection. a The tissues were stained with specific Abs against CD4 ( green ), CD25 ( white ), IL-17 ( red ), FOXP3 ( green ), p -STAT3–Y705 ( red ), and p -STAT3–S727 ( red ). Positive cells are shown in the bar graphs . b Spleen tissues were isolated from mice in each group at 14 weeks after curdlan injection. Splenocytes were stained with Abs against CD4–PerCP, IL-17–PE, and IL-10–FITC to determine the presence of Th17 and IL-10-expressing Treg cells. Data are expressed as the mean ± SEM of 3 independent experiments (* P

    Journal: Journal of Translational Medicine

    Article Title: Rebamipide prevents peripheral arthritis and intestinal inflammation by reciprocally regulating Th17/Treg cell imbalance in mice with curdlan-induced spondyloarthritis

    doi: 10.1186/s12967-016-0942-5

    Figure Lengend Snippet: Rebamipide reciprocally regulates Th17/Treg cell imbalance. Spleen tissues were isolated from mice in each group at 14 weeks after curdlan injection. a The tissues were stained with specific Abs against CD4 ( green ), CD25 ( white ), IL-17 ( red ), FOXP3 ( green ), p -STAT3–Y705 ( red ), and p -STAT3–S727 ( red ). Positive cells are shown in the bar graphs . b Spleen tissues were isolated from mice in each group at 14 weeks after curdlan injection. Splenocytes were stained with Abs against CD4–PerCP, IL-17–PE, and IL-10–FITC to determine the presence of Th17 and IL-10-expressing Treg cells. Data are expressed as the mean ± SEM of 3 independent experiments (* P

    Article Snippet: Cells were permeabilized and fixed with CytoFix (BD Biosciences, San Jose, CA, USA), as instructed by the manufacturer, and were stained with Abs against IL-17–PE and IL-10–FITC (eBioscience).

    Techniques: Isolation, Mouse Assay, Injection, Staining, Expressing

    Leishmania Ag–stimulated cytokine profiles in splenocytes culture supernatants from naive, LdCen-/- immunized (Imm), naive challenged (Naive Chal), and LdCen-/- immunized challenged (Imm Chal) young and aged mice. The 5-wk post immunized mice were challenged with virulent L . donovani for 4-wk and then mice were euthanized, splenocytes were isolated, plated aseptically (2×10 5 cells/well), and stimulated with Leishmania FTAg for 72h. Concentrations of pro-inflammatory cytokines IFN-γ (A), IL-12 (B) and TNF (C) anti-inflammatory cytokines IL-10 (D) and IL-4 (E) were measured in culture supernatants using the multiplex mouse cytokine kit as described in the Material and Methods section. Ratio of IFN-γ/IL-10 (F) and IFN-γ/IL-4 (G) were also determined. The data presented are representative of two independent experiments with similar results (n = 6). Mean and SEM of each group are shown. * p

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Live Attenuated Leishmania donovani Centrin Knock Out Parasites Generate Non-inferior Protective Immune Response in Aged Mice against Visceral Leishmaniasis

    doi: 10.1371/journal.pntd.0004963

    Figure Lengend Snippet: Leishmania Ag–stimulated cytokine profiles in splenocytes culture supernatants from naive, LdCen-/- immunized (Imm), naive challenged (Naive Chal), and LdCen-/- immunized challenged (Imm Chal) young and aged mice. The 5-wk post immunized mice were challenged with virulent L . donovani for 4-wk and then mice were euthanized, splenocytes were isolated, plated aseptically (2×10 5 cells/well), and stimulated with Leishmania FTAg for 72h. Concentrations of pro-inflammatory cytokines IFN-γ (A), IL-12 (B) and TNF (C) anti-inflammatory cytokines IL-10 (D) and IL-4 (E) were measured in culture supernatants using the multiplex mouse cytokine kit as described in the Material and Methods section. Ratio of IFN-γ/IL-10 (F) and IFN-γ/IL-4 (G) were also determined. The data presented are representative of two independent experiments with similar results (n = 6). Mean and SEM of each group are shown. * p

    Article Snippet: Culture supernatants were collected at 24h post infection to evaluate cytokine (IL-12, TNF and IL-6) production with the use of sandwich ELISA kit (ebioscience) and nitric oxide production by the Griess assay.

    Techniques: Mouse Assay, Isolation, Multiplex Assay

    Ag-specific intracellular cytokine secretion analysis of CD4 and CD8 T cells from LdCen-/- immunized and non-immunized young and aged mice after virulent L . donovani challenge. The 8- wk post-immunized or non-immunized young and aged mice were challenged for 4-wk with virulent L . donovani . Intracellular cytokine analysis was done as shown in Fig 6A and divided into seven distinct subpopulations. (A) Cytokine analysis of CD4 T cells from 8-wk post immunized and 4-wk post challenged mice. (B) Cytokine analysis of CD8 T cells of 8-wk post immunized and 4-wk post challenged mice. (C) IL-10 secreting CD4 T cells and (D) the ratio of IFN-γ to IL-10 producing CD4 T cells from spleens at the time of challenge [(naive and immunized (8W)] and after challenge [(naive-challenged and immune-challenged (8WI plus 4WPC)]. The data presented are representative of two experiments with similar results. Mean and SEM of six mice in each group are shown. * p

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Live Attenuated Leishmania donovani Centrin Knock Out Parasites Generate Non-inferior Protective Immune Response in Aged Mice against Visceral Leishmaniasis

    doi: 10.1371/journal.pntd.0004963

    Figure Lengend Snippet: Ag-specific intracellular cytokine secretion analysis of CD4 and CD8 T cells from LdCen-/- immunized and non-immunized young and aged mice after virulent L . donovani challenge. The 8- wk post-immunized or non-immunized young and aged mice were challenged for 4-wk with virulent L . donovani . Intracellular cytokine analysis was done as shown in Fig 6A and divided into seven distinct subpopulations. (A) Cytokine analysis of CD4 T cells from 8-wk post immunized and 4-wk post challenged mice. (B) Cytokine analysis of CD8 T cells of 8-wk post immunized and 4-wk post challenged mice. (C) IL-10 secreting CD4 T cells and (D) the ratio of IFN-γ to IL-10 producing CD4 T cells from spleens at the time of challenge [(naive and immunized (8W)] and after challenge [(naive-challenged and immune-challenged (8WI plus 4WPC)]. The data presented are representative of two experiments with similar results. Mean and SEM of six mice in each group are shown. * p

    Article Snippet: Culture supernatants were collected at 24h post infection to evaluate cytokine (IL-12, TNF and IL-6) production with the use of sandwich ELISA kit (ebioscience) and nitric oxide production by the Griess assay.

    Techniques: Mouse Assay

    Multiparameter flow cytometry based analysis for single, double, or triple cytokine–secreting CD44 Hi /CCR7 Low / CD4 or CD8 T cells from LdCen-/- immunized and non-immunized young and aged mice. (A) The common gating steps shown in this study. Spleen cells of 8-wk post immunized mice were stimulated with FTAg for 48h and stained with various Abs as described in Materials and Methods. Ag-experienced effector cells were gated and divided into seven distinct subpopulations, and the frequencies of the various subpopulations were calculated. (B) Cytokine analysis of CD4 T cells from naïve and 8-wk post-immunized mice. ( C ) Cytokine analysis of CD8 T cells from naïve and 8-wk post-immunized mice. The data presented are representative of two experiments with similar results. Mean and SEM of six in each group is shown. * p

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Live Attenuated Leishmania donovani Centrin Knock Out Parasites Generate Non-inferior Protective Immune Response in Aged Mice against Visceral Leishmaniasis

    doi: 10.1371/journal.pntd.0004963

    Figure Lengend Snippet: Multiparameter flow cytometry based analysis for single, double, or triple cytokine–secreting CD44 Hi /CCR7 Low / CD4 or CD8 T cells from LdCen-/- immunized and non-immunized young and aged mice. (A) The common gating steps shown in this study. Spleen cells of 8-wk post immunized mice were stimulated with FTAg for 48h and stained with various Abs as described in Materials and Methods. Ag-experienced effector cells were gated and divided into seven distinct subpopulations, and the frequencies of the various subpopulations were calculated. (B) Cytokine analysis of CD4 T cells from naïve and 8-wk post-immunized mice. ( C ) Cytokine analysis of CD8 T cells from naïve and 8-wk post-immunized mice. The data presented are representative of two experiments with similar results. Mean and SEM of six in each group is shown. * p

    Article Snippet: Culture supernatants were collected at 24h post infection to evaluate cytokine (IL-12, TNF and IL-6) production with the use of sandwich ELISA kit (ebioscience) and nitric oxide production by the Griess assay.

    Techniques: Flow Cytometry, Cytometry, Mouse Assay, Staining

    Analysis of young and aged mice derived dendritic cell function upon LdWT and LdCen-/- infection in vitro . BMDCs isolated from young and aged groups of mice were infected with LdWT or LdCen-/- stationary-phase promastigotes (5:1, parasite to DCs ratio) and intracellular parasite numbers were visualized by Giemsa staining and estimated microscopically at 6, 24, 48 and 72h post-infection. (A) Infection efficiency (% of infected cells) and (B) intracellular growth (parasites per infected cell) were recorded. To measure parasite load in these cultures, a minimum of 300 BMDCs were counted. In a separate experiment BMDCs were infected with parasites and stimulated with LPS (1 μg/ml) for 24h. Culture supernatants were collected to analyze IL-12 (C), TNF (D) IL-6 (E) production by ELISA and NO (F) production by the Griess assay. The data presented are representative of two independent experiments. T cell proliferation and cytokine production upon coculture of parasite-infected BMDCs with OVA-specific transgenic T cells. BMDCs obtained from young and aged BALB/c mice were pulsed with OVA peptide and were either left uninfected or infected with LdWT or LdCen-/- for 24h. CD4 + T cells were purified from age matched young and aged DO11.10 transgenic mice, stained with CFSE and co-cultured with parasite infected BMDCs for 5 days. (G) T cell proliferation was estimated by flow cytometry by studying CFSE dilution of gated CD4 + T cells and represented by the histogram. The staggered histogram overlay displays CD4 + T cell proliferation pattern as visualized by CFSE dilution by flow cytometry. Cell proliferation was done in triplicates and histograms representative of mean values were overlaid for figure. The black line on histogram over lay represents % proliferated cells gated in CD4 + T lymphocytes. (H) The bar diagram representing the quantitative CFSE cell proliferation. (I, J) Culture supernatants were collected at day 5 of coculture to assay IFN-γ and IL-10 by ELISA. (K) IFN-γ: IL-10 ratio was determined. The data represent the mean values ± SD of results from 3 independent experiments that all yielded similar results. * p

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Live Attenuated Leishmania donovani Centrin Knock Out Parasites Generate Non-inferior Protective Immune Response in Aged Mice against Visceral Leishmaniasis

    doi: 10.1371/journal.pntd.0004963

    Figure Lengend Snippet: Analysis of young and aged mice derived dendritic cell function upon LdWT and LdCen-/- infection in vitro . BMDCs isolated from young and aged groups of mice were infected with LdWT or LdCen-/- stationary-phase promastigotes (5:1, parasite to DCs ratio) and intracellular parasite numbers were visualized by Giemsa staining and estimated microscopically at 6, 24, 48 and 72h post-infection. (A) Infection efficiency (% of infected cells) and (B) intracellular growth (parasites per infected cell) were recorded. To measure parasite load in these cultures, a minimum of 300 BMDCs were counted. In a separate experiment BMDCs were infected with parasites and stimulated with LPS (1 μg/ml) for 24h. Culture supernatants were collected to analyze IL-12 (C), TNF (D) IL-6 (E) production by ELISA and NO (F) production by the Griess assay. The data presented are representative of two independent experiments. T cell proliferation and cytokine production upon coculture of parasite-infected BMDCs with OVA-specific transgenic T cells. BMDCs obtained from young and aged BALB/c mice were pulsed with OVA peptide and were either left uninfected or infected with LdWT or LdCen-/- for 24h. CD4 + T cells were purified from age matched young and aged DO11.10 transgenic mice, stained with CFSE and co-cultured with parasite infected BMDCs for 5 days. (G) T cell proliferation was estimated by flow cytometry by studying CFSE dilution of gated CD4 + T cells and represented by the histogram. The staggered histogram overlay displays CD4 + T cell proliferation pattern as visualized by CFSE dilution by flow cytometry. Cell proliferation was done in triplicates and histograms representative of mean values were overlaid for figure. The black line on histogram over lay represents % proliferated cells gated in CD4 + T lymphocytes. (H) The bar diagram representing the quantitative CFSE cell proliferation. (I, J) Culture supernatants were collected at day 5 of coculture to assay IFN-γ and IL-10 by ELISA. (K) IFN-γ: IL-10 ratio was determined. The data represent the mean values ± SD of results from 3 independent experiments that all yielded similar results. * p

    Article Snippet: Culture supernatants were collected at 24h post infection to evaluate cytokine (IL-12, TNF and IL-6) production with the use of sandwich ELISA kit (ebioscience) and nitric oxide production by the Griess assay.

    Techniques: Mouse Assay, Derivative Assay, Cell Function Assay, Infection, In Vitro, Isolation, Staining, Enzyme-linked Immunosorbent Assay, Griess Assay, Transgenic Assay, Purification, Cell Culture, Flow Cytometry, Cytometry

    Indoleamine 2,3 dioxygenase-1 (IDO1) influences the gene expression of cytokines and transcription factors. Relative expression of mRNA for aryl hydrocarbon receptor (AhR), IFN-γ, TNF-α, IL-6, RORC, Tbet, GATA3, FoxP3, IL-10, TGF-β, IL-17, and IL-22 in whole lung cells of wild-type and IDO1 −/− mice after 10 weeks of Paracoccidioides brasiliensis infection. The level of gene transcription was determined by real-time PCR. Bars show mean ± SEM from at least four mice per group and are representative of three independent experiments (* p

    Journal: Frontiers in Immunology

    Article Title: The IDO–AhR Axis Controls Th17/Treg Immunity in a Pulmonary Model of Fungal Infection

    doi: 10.3389/fimmu.2017.00880

    Figure Lengend Snippet: Indoleamine 2,3 dioxygenase-1 (IDO1) influences the gene expression of cytokines and transcription factors. Relative expression of mRNA for aryl hydrocarbon receptor (AhR), IFN-γ, TNF-α, IL-6, RORC, Tbet, GATA3, FoxP3, IL-10, TGF-β, IL-17, and IL-22 in whole lung cells of wild-type and IDO1 −/− mice after 10 weeks of Paracoccidioides brasiliensis infection. The level of gene transcription was determined by real-time PCR. Bars show mean ± SEM from at least four mice per group and are representative of three independent experiments (* p

    Article Snippet: The levels of IL-1β, IL-2, IL-4, IL-6, IL-10, IL-12, IL-17, IL-22, IL-27, IL-35, IL-23, TNF-α, IFN-γ, and TGF-β were measured by capture enzyme-linked immunosorbent assay (ELISA) with antibody pairs purchased from eBioscience or PBL.

    Techniques: Expressing, Mouse Assay, Infection, Real-time Polymerase Chain Reaction

    Indoleamine 2,3 dioxygenase-1 (IDO1) expression controls the presence of intracellular cytokines in pulmonary CD11b + and CD11c + cells. The intracellular cytokines were determined in CD11b + (A) and CD11c + (B) lung infiltrating leukocytes of wild-type and IDO1 −/− C57BL/6 mice after 96 h, 2, and 10 weeks of infection with 1 × 10 6 Paracoccidioides brasiliensis yeasts. The lung cells were obtained as described in Section “ Materials and Methods ” and labeled with antibodies conjugated to different fluorochromes. The lung infiltrating leukocytes were gated by FSC/SSC analysis. The cells were gated for CD11b + or CD11c + expression and then for the presence of cytokines IL-12, TNF-α, IL-1β, IL-10, IL-6, and TGF-β. One hundred thousand cells were acquired on FACS CANTO II and subsequently analyzed by FlowJo software. Data are expressed as means ± SEM and are representative of three independent experiments using five mice of each mouse strain per group (* p

    Journal: Frontiers in Immunology

    Article Title: The IDO–AhR Axis Controls Th17/Treg Immunity in a Pulmonary Model of Fungal Infection

    doi: 10.3389/fimmu.2017.00880

    Figure Lengend Snippet: Indoleamine 2,3 dioxygenase-1 (IDO1) expression controls the presence of intracellular cytokines in pulmonary CD11b + and CD11c + cells. The intracellular cytokines were determined in CD11b + (A) and CD11c + (B) lung infiltrating leukocytes of wild-type and IDO1 −/− C57BL/6 mice after 96 h, 2, and 10 weeks of infection with 1 × 10 6 Paracoccidioides brasiliensis yeasts. The lung cells were obtained as described in Section “ Materials and Methods ” and labeled with antibodies conjugated to different fluorochromes. The lung infiltrating leukocytes were gated by FSC/SSC analysis. The cells were gated for CD11b + or CD11c + expression and then for the presence of cytokines IL-12, TNF-α, IL-1β, IL-10, IL-6, and TGF-β. One hundred thousand cells were acquired on FACS CANTO II and subsequently analyzed by FlowJo software. Data are expressed as means ± SEM and are representative of three independent experiments using five mice of each mouse strain per group (* p

    Article Snippet: The levels of IL-1β, IL-2, IL-4, IL-6, IL-10, IL-12, IL-17, IL-22, IL-27, IL-35, IL-23, TNF-α, IFN-γ, and TGF-β were measured by capture enzyme-linked immunosorbent assay (ELISA) with antibody pairs purchased from eBioscience or PBL.

    Techniques: Expressing, Mouse Assay, Infection, Labeling, FACS, Software

    IL-6 downregulation in SOD1-G93A mouse microglia. ( a ) Semiquantitative RT-PCR using specific primers for IL-6 and β -actin mRNAs, on total RNA from nt and SOD1-G93A microglia. β -actin was used for normalization. ( b ) Western blotting with anti-IL-6 antibody on total lysates from nt and SOD1-G93A microglia. β -actin was used for protein normalization. ( c ) IL-6 levels in the culture media of microglia from nt and G93A mice, after 6 h incubation in fresh media, as assessed by ELISA

    Journal: Cell Death & Disease

    Article Title: Dysregulated microRNAs in amyotrophic lateral sclerosis microglia modulate genes linked to neuroinflammation

    doi: 10.1038/cddis.2013.491

    Figure Lengend Snippet: IL-6 downregulation in SOD1-G93A mouse microglia. ( a ) Semiquantitative RT-PCR using specific primers for IL-6 and β -actin mRNAs, on total RNA from nt and SOD1-G93A microglia. β -actin was used for normalization. ( b ) Western blotting with anti-IL-6 antibody on total lysates from nt and SOD1-G93A microglia. β -actin was used for protein normalization. ( c ) IL-6 levels in the culture media of microglia from nt and G93A mice, after 6 h incubation in fresh media, as assessed by ELISA

    Article Snippet: ELISA Microglia were incubated for 6 h in fresh media without serum, and mouse IL-6 expression was measured in the supernatants using a commercial mouse IL-6 ELISA kit (Invitrogen), according to the manufacturer's instructions.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Mouse Assay, Incubation, Enzyme-linked Immunosorbent Assay