anti il17 antibody  (Thermo Fisher)


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    Name:
    anti IL 17
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    PY-10007
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    Thermo Fisher anti il17 antibody
    IL-17 production during Salmonella infection in WT and Crtam −/− mice. Immune cells from the gut of infected mice were isolated, then stimulated with PMA and ionomycin in the presence of brefeldin-A. After 6 hours of stimulation, cells were stained for intracellular IL-17, then analyzed by flow cytometry. ( a ) Representative contour plots of CD3 + CD4 + IL-17 + and CD3 + CD4 + CD8 + IL-17 + cells obtained from the gut of S . Typhimurium-infected WT ( n =12) or Crtam −/− ( n =11) mice are shown. ( b ) Frequency of IL-17-producing cells per million live cells was calculated for each T cell subset. Data shown comprise three independent experiments. ( c, d ) Cells from the small intestine were fractionated into ( c ) intraepithelial lymphocytes (IEL) and ( d ) lamina propria lymphocytes (LPL), then treated as mentioned above. Frequency of IL-17-producing cells per million live cells was calculated for each indicated T cell subset. ( b, c, d ) Each black circle (WT) or white square ( Crtam −/− ) represents a mouse, and bars represent the average of each group. ( e ) Relative expression levels (qPCR) of Il1b, Il6, Il23, and Tgfb in the cecum of WT (black bars, n =8) or Crtam −/− (white bars, n =11) mice at 96h post- S . Typhimurium infection, compared to uninfected controls. ( f ) Relative expression levels (qPCR) of <t>Il17,</t> Il22, Lcn2, and Cxcl1 in the cecum of WT (black bars, n =10) or Crtam −/− (white bars, n =8) mice at 96h post- S . Typhimurium infection, compared to uninfected controls. Expression of Actb was used as a housekeeping control. Data represent the mean ± standard error. Data shown comprise three independent experiments. n.s. = not significant. A significant difference (t-test) is indicated by * ( P ≤ 0.05) or ** ( P ≤ 0.01).

    https://www.bioz.com/result/anti il17 antibody/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti il17 antibody - by Bioz Stars, 2021-06
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    Images

    1) Product Images from "CRTAM shapes the gut microbiota and enhances the severity of infection"

    Article Title: CRTAM shapes the gut microbiota and enhances the severity of infection

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

    doi: 10.4049/jimmunol.1800890

    IL-17 production during Salmonella infection in WT and Crtam −/− mice. Immune cells from the gut of infected mice were isolated, then stimulated with PMA and ionomycin in the presence of brefeldin-A. After 6 hours of stimulation, cells were stained for intracellular IL-17, then analyzed by flow cytometry. ( a ) Representative contour plots of CD3 + CD4 + IL-17 + and CD3 + CD4 + CD8 + IL-17 + cells obtained from the gut of S . Typhimurium-infected WT ( n =12) or Crtam −/− ( n =11) mice are shown. ( b ) Frequency of IL-17-producing cells per million live cells was calculated for each T cell subset. Data shown comprise three independent experiments. ( c, d ) Cells from the small intestine were fractionated into ( c ) intraepithelial lymphocytes (IEL) and ( d ) lamina propria lymphocytes (LPL), then treated as mentioned above. Frequency of IL-17-producing cells per million live cells was calculated for each indicated T cell subset. ( b, c, d ) Each black circle (WT) or white square ( Crtam −/− ) represents a mouse, and bars represent the average of each group. ( e ) Relative expression levels (qPCR) of Il1b, Il6, Il23, and Tgfb in the cecum of WT (black bars, n =8) or Crtam −/− (white bars, n =11) mice at 96h post- S . Typhimurium infection, compared to uninfected controls. ( f ) Relative expression levels (qPCR) of Il17, Il22, Lcn2, and Cxcl1 in the cecum of WT (black bars, n =10) or Crtam −/− (white bars, n =8) mice at 96h post- S . Typhimurium infection, compared to uninfected controls. Expression of Actb was used as a housekeeping control. Data represent the mean ± standard error. Data shown comprise three independent experiments. n.s. = not significant. A significant difference (t-test) is indicated by * ( P ≤ 0.05) or ** ( P ≤ 0.01).
    Figure Legend Snippet: IL-17 production during Salmonella infection in WT and Crtam −/− mice. Immune cells from the gut of infected mice were isolated, then stimulated with PMA and ionomycin in the presence of brefeldin-A. After 6 hours of stimulation, cells were stained for intracellular IL-17, then analyzed by flow cytometry. ( a ) Representative contour plots of CD3 + CD4 + IL-17 + and CD3 + CD4 + CD8 + IL-17 + cells obtained from the gut of S . Typhimurium-infected WT ( n =12) or Crtam −/− ( n =11) mice are shown. ( b ) Frequency of IL-17-producing cells per million live cells was calculated for each T cell subset. Data shown comprise three independent experiments. ( c, d ) Cells from the small intestine were fractionated into ( c ) intraepithelial lymphocytes (IEL) and ( d ) lamina propria lymphocytes (LPL), then treated as mentioned above. Frequency of IL-17-producing cells per million live cells was calculated for each indicated T cell subset. ( b, c, d ) Each black circle (WT) or white square ( Crtam −/− ) represents a mouse, and bars represent the average of each group. ( e ) Relative expression levels (qPCR) of Il1b, Il6, Il23, and Tgfb in the cecum of WT (black bars, n =8) or Crtam −/− (white bars, n =11) mice at 96h post- S . Typhimurium infection, compared to uninfected controls. ( f ) Relative expression levels (qPCR) of Il17, Il22, Lcn2, and Cxcl1 in the cecum of WT (black bars, n =10) or Crtam −/− (white bars, n =8) mice at 96h post- S . Typhimurium infection, compared to uninfected controls. Expression of Actb was used as a housekeeping control. Data represent the mean ± standard error. Data shown comprise three independent experiments. n.s. = not significant. A significant difference (t-test) is indicated by * ( P ≤ 0.05) or ** ( P ≤ 0.01).

    Techniques Used: Infection, Mouse Assay, Isolation, Staining, Flow Cytometry, Expressing, Real-time Polymerase Chain Reaction, T-Test

    2) Product Images from "Th17 Pathway As a Target for Multipotent Stromal Cell Therapy in Dogs: Implications for Translational Research"

    Article Title: Th17 Pathway As a Target for Multipotent Stromal Cell Therapy in Dogs: Implications for Translational Research

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0148568

    Th17 detection strategy and baseline levels in healthy dogs. Panel A depicts the gating strategy that was employed to detect circulating Th17 cells in dogs. Peripheral blood mononuclear cells were isolated, stimulated with PMA and ionomycin and stained with CD3, CD4, IL17 and a viability dye as described in the methods. Viable cells were gated and intracellular IL17 was detected within Th cells (i.e. CD3+/CD4+), non-Th cells (primarily CD8 T cells, CD3+/CD4-) and within non T cells (primarily B cells, CD3-). The percentages of the various lymphocyte subsets that were IL17 positive (B) and the MFI of IL17 within Th17 cells (i.e. CD3+/CD4+) as compared with CD4- T cells (C) are depicted. Note that while CD4+ and CD4- T cells had similar proportion of IL17 positive cells (B), CD4+ T cells had higher MFI compared with CD4- T cells (C), suggesting greater capacity to produce IL17 upon stimulation. Data are represented as mean +/- standard deviation, n = 5. (* P
    Figure Legend Snippet: Th17 detection strategy and baseline levels in healthy dogs. Panel A depicts the gating strategy that was employed to detect circulating Th17 cells in dogs. Peripheral blood mononuclear cells were isolated, stimulated with PMA and ionomycin and stained with CD3, CD4, IL17 and a viability dye as described in the methods. Viable cells were gated and intracellular IL17 was detected within Th cells (i.e. CD3+/CD4+), non-Th cells (primarily CD8 T cells, CD3+/CD4-) and within non T cells (primarily B cells, CD3-). The percentages of the various lymphocyte subsets that were IL17 positive (B) and the MFI of IL17 within Th17 cells (i.e. CD3+/CD4+) as compared with CD4- T cells (C) are depicted. Note that while CD4+ and CD4- T cells had similar proportion of IL17 positive cells (B), CD4+ T cells had higher MFI compared with CD4- T cells (C), suggesting greater capacity to produce IL17 upon stimulation. Data are represented as mean +/- standard deviation, n = 5. (* P

    Techniques Used: Isolation, Staining, Standard Deviation

    IL17 and CD3 dual detection in idiopathic chronic inflammatory lesion in dogs. FFPE tissues from dogs with naturally occurring diseases were identified, stained and scanned as described in the Material and Methods section. Each panel (A-F) consists of a low power (objective X4) image of an H E stained slide and a high power (objective X63) image of the same slide stained with CD3 (green), IL17 (red) and DAPI (blue). Presented are representative images from the duodenum (A) and mesenteric lymph node (B) of dogs (n = 5) with inflammatory bowel disease, skin (C) of a dog with chronic idiopathic dermatitis, gingiva (D) of dogs (n = 5) with chronic idiopathic gingivitis, cerebrum (E) of a dog (n = 1) with necrotizing meningoencephalitis and nasal mucosa (E) of dogs (n = 3) with chronic rhinitis. CD3 positive (i.e. green) and IL17 positive (i.e. red) cells are present in all of the represented lesions. Low numbers of double positive cells (indicated by white arrows) are present while the majority of the IL17 positive cells are CD3 negative.
    Figure Legend Snippet: IL17 and CD3 dual detection in idiopathic chronic inflammatory lesion in dogs. FFPE tissues from dogs with naturally occurring diseases were identified, stained and scanned as described in the Material and Methods section. Each panel (A-F) consists of a low power (objective X4) image of an H E stained slide and a high power (objective X63) image of the same slide stained with CD3 (green), IL17 (red) and DAPI (blue). Presented are representative images from the duodenum (A) and mesenteric lymph node (B) of dogs (n = 5) with inflammatory bowel disease, skin (C) of a dog with chronic idiopathic dermatitis, gingiva (D) of dogs (n = 5) with chronic idiopathic gingivitis, cerebrum (E) of a dog (n = 1) with necrotizing meningoencephalitis and nasal mucosa (E) of dogs (n = 3) with chronic rhinitis. CD3 positive (i.e. green) and IL17 positive (i.e. red) cells are present in all of the represented lesions. Low numbers of double positive cells (indicated by white arrows) are present while the majority of the IL17 positive cells are CD3 negative.

    Techniques Used: Formalin-fixed Paraffin-Embedded, Staining

    Induction of Th17 polarization in canine Th cells in vitro. Peripheral blood derived CD4+ T cells from healthy dogs (n = 7) were isolated and stimulated as described in the Material and Methods section. Th17 polarization was determined by flow cytometric detection of intracellular IL17 (A), secreted IL17 via ELISA (B) and gene transcription via qRT-PCR (C). Th17 detection by flow cytometry was strongly correlated with ELISA detection of secreted IL17 and both had reached a maximum at 7 days post stimulation (P
    Figure Legend Snippet: Induction of Th17 polarization in canine Th cells in vitro. Peripheral blood derived CD4+ T cells from healthy dogs (n = 7) were isolated and stimulated as described in the Material and Methods section. Th17 polarization was determined by flow cytometric detection of intracellular IL17 (A), secreted IL17 via ELISA (B) and gene transcription via qRT-PCR (C). Th17 detection by flow cytometry was strongly correlated with ELISA detection of secreted IL17 and both had reached a maximum at 7 days post stimulation (P

    Techniques Used: In Vitro, Derivative Assay, Isolation, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Flow Cytometry

    Related Articles

    Staining:

    Article Title: CRTAM shapes the gut microbiota and enhances the severity of infection
    Article Snippet: Briefly, cells were blocked with a CD16/32 antibody (eBioscience), stained with the viability dye eFluor780 (eBioscience), then extracellularly stained using the following monoclonal antibodies: CD45 (clone 30-F11), CD3 (clone 17A2), CD4 (clone RM4–5), CD8α (clone 53–6.7) – each from BioLegend; TCR γδ (clone eBio-GL3) and CD25 (clone PC61.5) – each from eBioscience. .. After surface staining, cells were fixed and permeabilized according to the manufacturer’s instructions (“Fix and Perm” kit, eBioscience), then stained intracellularly with anti-IL17 antibody (clone TC11–18H10.1), anti-IFNγ antibody (clone XMG1.2), and anti-IL-10 antibody (clone JES5–16-E3) from eBioscience. .. Cells were analyzed on an LSRII flow cytometer (BD Biosciences, San Jose, CA) and analyzed with FlowJo software (TreeStar, Ashland, OR).

    Article Title: Ginseng Berry Extract Attenuates Dextran Sodium Sulfate-Induced Acute and Chronic Colitis
    Article Snippet: Ex Vivo Stimulation of LP Cells and Intracellular Cytokine Staining As described in detail previously [ ], single cell suspensions from the LP were stimulated with phorbol 12-myristate 13-acetate (PMA) (50 ng/mL) and ionomycin (1 μM; both from Calbiochem) for 4 h, with the addition of monensin (eBioscience, San Diego, CA, USA) in the final 2 h. Cells were then stained for NK1.1, Ly-6G, TCR-β, and TCR-γδ to determine the phenotypes of the infiltrating leukocytes. .. For intracellular cytokine staining, LP cells were surface stained with CD4, CD8, and TCR-β, and then fixed and permeabilized with Cytofix/Cytoperm buffer (eBioscience, San Diego, CA, USA), followed by incubation with anti-IL-17 and anti-IFN-γ antibodies in Perm/Wash buffer (eBioscience, San Diego, CA, USA) for 30 min. Control staining with isotype control IgGs was performed in all experiments. .. Intestinal Macrophage and DC Analysis As described in detail previously [ ], single cell suspensions from the colon were incubated for 30 min with the following fluorescence-conjugated monoclonal antibodies (mAbs): anti-CD45, anti-CD11c, anti-MHC class II, anti-CD103, and anti-F4/80.

    Article Title: IL-17RA Is Required for CCL2 Expression, Macrophage Recruitment, and Emphysema in Response to Cigarette Smoke
    Article Snippet: Cells were surface stained for 15–30 min at 4°C with anti-CD4 mAb (RM45; BD Pharmingen, San Diego, CA) in PBS supplemented with 1% BSA and 0.2% sodium azide. .. Cells were then fixed and permeabilized with Cytofix/Cytoperm (BD Pharmingen, San Diego, CA) and stained intracellularly with anti-IL-17, anti-IL-22 and anti-IFN-γ (XMG1.2) (eBioscience, San Diego, CA). .. Samples were acquired on a FACSAria or LSR-II flow cytometer and data analysis was conducted using FlowJo software (Treestar).

    Article Title: Role of IL-17A on Resolution of Pulmonary C. neoformans Infection
    Article Snippet: For intracellular staining, cells remained in fixation buffer for 10 min at room temperature. .. While permeabilized, the cells were intracellularly stained with anti-IL-17A (eBioscience Inc.) and/or anti-Fox3P (regulatory T cell) (eBioscience Inc.) for 30 min at 4°C. .. Cells were then washed 3 times with 0.1% saponin and then resuspended in fixation buffer before flow cytometry was performed.

    Article Title: Differential Frequency of CD8+ T Cell Subsets in Multiple Sclerosis Patients with Various Clinical Patterns
    Article Snippet: After surface staining, the cells were fixed and permeabilized using LEUCOPERM™ (BIO-RAD, BUF09B, US) according to the manufacturer’s instructions. .. The cells were stained with anti-IFN-γ-FITC, anti-TNF-α-PE-Cy™ 7, anti-IL-10-eFluor® 450, anti-IL-17-PE, anti-IL-21-eFluor® 660, anti-IL-22-PerCP-eFluor® 710 (eBioscience, US) and anti- IL-4-APC-Cy™ 7 (BioLegend, US). .. Enzyme-Linked Immunosorbent Assay (ELISA) For investigation of cytokines related to Tc17 development, sera of patients and controls were collected from clot tubes by centrifugation at 1500rpm for 10 minutes at 4°C.

    Article Title: Immunological benefits by ginseng through reciprocal regulation of Th17 and Treg cells during cyclosporine-induced immunosuppression
    Article Snippet: 2.4 Flow cytometry Expression of cytokines and transcription factors was assessed by intracellular staining. .. The following antibodies were used for intracellular staining of mouse cells: anti-CD4- peridinin chlorophyll or -fluorescein isothiocyanate, anti-CD25-eFluor 450, anti-IL-17-phycoerythrin (PE), anti-Foxp3-allophycocyanin (APC), anti-IFNγ-peridinin chlorophyll-cyanine 5.5, and anti-IL-4-PE-cyanine 7 (all from eBioscience, San Diego, CA, USA). .. Cells were stimulated for 4 h with phorbol 12-myristate 13-acetate (Sigma) and ionomycin (Sigma) with the addition of GolgiStop (BD Bioscience).

    Article Title: IL-22 promotes allergic airway inflammation in epicutaneously sensitized mice
    Article Snippet: For Intracellular staining, lung cells were stimulated with PMA (50 ng/ml) and ionomycin (500 ng/ml) in presence of Golgi-stop and Golgi-plug for 4 hours in IMDM media, followed by surface staining for CD3, CD4, CD8, γδ TCR and CD90. .. Cells were then fixed and permeabilized using BD Cytofix/Cytoperm Kit (BD Biosciences) and stained with anti-IL-22 (Biolegend), anti-TNFα (Biolegend), anti-IL-17A (eBioscience) or anti-IFNγ (eBioscience) antibodies.. .. Finally, cells were washed and analyzed on LSRFortessa (BD Biosciences).

    Incubation:

    Article Title: Ginseng Berry Extract Attenuates Dextran Sodium Sulfate-Induced Acute and Chronic Colitis
    Article Snippet: Ex Vivo Stimulation of LP Cells and Intracellular Cytokine Staining As described in detail previously [ ], single cell suspensions from the LP were stimulated with phorbol 12-myristate 13-acetate (PMA) (50 ng/mL) and ionomycin (1 μM; both from Calbiochem) for 4 h, with the addition of monensin (eBioscience, San Diego, CA, USA) in the final 2 h. Cells were then stained for NK1.1, Ly-6G, TCR-β, and TCR-γδ to determine the phenotypes of the infiltrating leukocytes. .. For intracellular cytokine staining, LP cells were surface stained with CD4, CD8, and TCR-β, and then fixed and permeabilized with Cytofix/Cytoperm buffer (eBioscience, San Diego, CA, USA), followed by incubation with anti-IL-17 and anti-IFN-γ antibodies in Perm/Wash buffer (eBioscience, San Diego, CA, USA) for 30 min. Control staining with isotype control IgGs was performed in all experiments. .. Intestinal Macrophage and DC Analysis As described in detail previously [ ], single cell suspensions from the colon were incubated for 30 min with the following fluorescence-conjugated monoclonal antibodies (mAbs): anti-CD45, anti-CD11c, anti-MHC class II, anti-CD103, and anti-F4/80.

    other:

    Article Title: NKT Cell Stimulation with ?-Galactosylceramide Results in a Block of Th17 Differentiation after Intranasal Immunization in Mice
    Article Snippet: Antibodies The following antibodies have been used: anti-IL-17A-APC, anti-IL-17A-PE, anti-IL-17F-APC, anti-IL-22-PerCP-eF710, anti-CD4-PE-Cy7, anti-CD44-APC, anti-NK1.1-APC, anti-IL-4, anti-IL-10 and anti-IFNγ from eBioscience; anti-CD62L-FITC, anti-CD8-PE, anti-B220-PE, anti-CD11c-PE, anti-CD11b-PE, anti-DX5-PE from BD Bioscience.

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    Thermo Fisher anti il 17a
    EC sensitization elicits a systemic IL-22 response and an antigen-specific IL-22 response in the lungs A-B. IL-22 secretion by OVA stimulated splenocytes (A) and IL-22 serum levels (B). C,D . Il22 mRNA expression in the lungs (C), and IL-22 secretion by OVA stimulated lung cells (D). E. Representative FACS analysis and quantitation of intracelluar expression of IL-22 + cells among CD3 + CD4 + T cells and of <t>IL-17A</t> + and TNFα + cells among CD3 + CD4 + IL-22 + cells in the lung. Mice were EC sensitized with OVA or saline in A and B, followed by i.n. challenged with OVA in C-F. Bars represent mean±SEM (n=5–10 per group). *p
    Anti Il 17a, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti il 17a/product/Thermo Fisher
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    93
    Thermo Fisher anti il 17 fitc
    FACS analyzed the CD3 + CD8 − <t>IL-17</t> + cell ratio in RA patient The PBMCs were isolated by standard Ficoll-Hypaque density centrifugation. The cells were stained by anti-CD3-PE-cy5, <t>anti-CD8-FITC,</t> and anti-IL-17-PE. (a) First figure presented CD3 + CD8 − T cells were considered CD4 + T cells in region RL, and the other three were presented healthy controls. (b) Representative IL-17 expression in CD3 + CD8 − T subsets from RA patients in inactive phase. (c) Representative IL-17 expression in CD3 + CD8 − T subsets from RA patients in active phase. (d) The results were shown as means ± SD. *P
    Anti Il 17 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
    https://www.bioz.com/result/anti il 17 fitc/product/Thermo Fisher
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    91
    Thermo Fisher pe anti mouse il 17 antibody
    <t>IFN-γ/IL-17</t> double knockout mice did not develop arthritis. (a) Comparison of arthritis score during 10 weeks after 1 st immunization. (b) Serum was collected at 5 th week after CIA induction, the level of anti CII IgG, IgG1, IgG2 was determined by ELISA. (c) Joint histology of CIA (IFN-γ KO, n = 14) and CIA (IFN-γIL-17 DKO, n = 14) at 10 th week after 1 st immunization. Joint tissue sections were stained with Hematoxylin and Eosin, Toluidine Blue, Safranin O. (d) Immunohistochemistry. RANKL or RANK positive cells were not seen in CIA (IFN-γ/IL-17 DKO) joint section. Original magnification,200x. Mann-Whitney U test (b) used. Values are presented as the mean ± standard deviation. *, p
    Pe Anti Mouse Il 17 Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher pe conjugated anti il 17 antibodies
    In vivo neutralization of interleukin <t>(IL)-17</t> does not protect against diabetes transferred by diabetic splenocytes. (a) Splenocytes from 15-week-old female diabetic or age- and sex-matched non-diabetic non-obese diabetic (NOD) mice were stimulated in
    Pe Conjugated Anti Il 17 Antibodies, 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


    EC sensitization elicits a systemic IL-22 response and an antigen-specific IL-22 response in the lungs A-B. IL-22 secretion by OVA stimulated splenocytes (A) and IL-22 serum levels (B). C,D . Il22 mRNA expression in the lungs (C), and IL-22 secretion by OVA stimulated lung cells (D). E. Representative FACS analysis and quantitation of intracelluar expression of IL-22 + cells among CD3 + CD4 + T cells and of IL-17A + and TNFα + cells among CD3 + CD4 + IL-22 + cells in the lung. Mice were EC sensitized with OVA or saline in A and B, followed by i.n. challenged with OVA in C-F. Bars represent mean±SEM (n=5–10 per group). *p

    Journal: The Journal of allergy and clinical immunology

    Article Title: IL-22 promotes allergic airway inflammation in epicutaneously sensitized mice

    doi: 10.1016/j.jaci.2018.05.032

    Figure Lengend Snippet: EC sensitization elicits a systemic IL-22 response and an antigen-specific IL-22 response in the lungs A-B. IL-22 secretion by OVA stimulated splenocytes (A) and IL-22 serum levels (B). C,D . Il22 mRNA expression in the lungs (C), and IL-22 secretion by OVA stimulated lung cells (D). E. Representative FACS analysis and quantitation of intracelluar expression of IL-22 + cells among CD3 + CD4 + T cells and of IL-17A + and TNFα + cells among CD3 + CD4 + IL-22 + cells in the lung. Mice were EC sensitized with OVA or saline in A and B, followed by i.n. challenged with OVA in C-F. Bars represent mean±SEM (n=5–10 per group). *p

    Article Snippet: Cells were then fixed and permeabilized using BD Cytofix/Cytoperm Kit (BD Biosciences) and stained with anti-IL-22 (Biolegend), anti-TNFα (Biolegend), anti-IL-17A (eBioscience) or anti-IFNγ (eBioscience) antibodies..

    Techniques: Expressing, FACS, Quantitation Assay, Mouse Assay

    FACS analyzed the CD3 + CD8 − IL-17 + cell ratio in RA patient The PBMCs were isolated by standard Ficoll-Hypaque density centrifugation. The cells were stained by anti-CD3-PE-cy5, anti-CD8-FITC, and anti-IL-17-PE. (a) First figure presented CD3 + CD8 − T cells were considered CD4 + T cells in region RL, and the other three were presented healthy controls. (b) Representative IL-17 expression in CD3 + CD8 − T subsets from RA patients in inactive phase. (c) Representative IL-17 expression in CD3 + CD8 − T subsets from RA patients in active phase. (d) The results were shown as means ± SD. *P

    Journal: Clinical and Developmental Immunology

    Article Title: Enhanced HMGB1 Expression May Contribute to Th17 Cells Activation in Rheumatoid Arthritis

    doi: 10.1155/2012/295081

    Figure Lengend Snippet: FACS analyzed the CD3 + CD8 − IL-17 + cell ratio in RA patient The PBMCs were isolated by standard Ficoll-Hypaque density centrifugation. The cells were stained by anti-CD3-PE-cy5, anti-CD8-FITC, and anti-IL-17-PE. (a) First figure presented CD3 + CD8 − T cells were considered CD4 + T cells in region RL, and the other three were presented healthy controls. (b) Representative IL-17 expression in CD3 + CD8 − T subsets from RA patients in inactive phase. (c) Representative IL-17 expression in CD3 + CD8 − T subsets from RA patients in active phase. (d) The results were shown as means ± SD. *P

    Article Snippet: 1 × 106 CD4+ T cells from the spleen of mice were stained with anti-IL-17-FITC (eBioscience).

    Techniques: FACS, Isolation, Centrifugation, Staining, Expressing

    IFN-γ/IL-17 double knockout mice did not develop arthritis. (a) Comparison of arthritis score during 10 weeks after 1 st immunization. (b) Serum was collected at 5 th week after CIA induction, the level of anti CII IgG, IgG1, IgG2 was determined by ELISA. (c) Joint histology of CIA (IFN-γ KO, n = 14) and CIA (IFN-γIL-17 DKO, n = 14) at 10 th week after 1 st immunization. Joint tissue sections were stained with Hematoxylin and Eosin, Toluidine Blue, Safranin O. (d) Immunohistochemistry. RANKL or RANK positive cells were not seen in CIA (IFN-γ/IL-17 DKO) joint section. Original magnification,200x. Mann-Whitney U test (b) used. Values are presented as the mean ± standard deviation. *, p

    Journal: PLoS ONE

    Article Title: Interferon Gamma Suppresses Collagen-Induced Arthritis by Regulation of Th17 through the Induction of Indoleamine-2,3-Deoxygenase

    doi: 10.1371/journal.pone.0060900

    Figure Lengend Snippet: IFN-γ/IL-17 double knockout mice did not develop arthritis. (a) Comparison of arthritis score during 10 weeks after 1 st immunization. (b) Serum was collected at 5 th week after CIA induction, the level of anti CII IgG, IgG1, IgG2 was determined by ELISA. (c) Joint histology of CIA (IFN-γ KO, n = 14) and CIA (IFN-γIL-17 DKO, n = 14) at 10 th week after 1 st immunization. Joint tissue sections were stained with Hematoxylin and Eosin, Toluidine Blue, Safranin O. (d) Immunohistochemistry. RANKL or RANK positive cells were not seen in CIA (IFN-γ/IL-17 DKO) joint section. Original magnification,200x. Mann-Whitney U test (b) used. Values are presented as the mean ± standard deviation. *, p

    Article Snippet: For intracellular FACS staining, cells were fixed with Cytofix/cytoperm solution (BD) followed by washing with permeablization buffer (BD) and then stained with PE anti mouse IL-17 antibody (ebioscience).

    Techniques: Double Knockout, Mouse Assay, Enzyme-linked Immunosorbent Assay, Staining, Immunohistochemistry, MANN-WHITNEY, Standard Deviation

    Suppression of IL-17 by IFN- γ is associated with IDO. (a) CD4 + T cells isolated from wild type (WT) C57BL/6 (B6) mice and IFN-γ knock out (KO) B6 mice. The cells were cultured in Th17-polarizing conditions (anti CD3 mAb, anti CD28 mAb, anti IFN-γAb, anti IL-4 Ab, TGF-β, IL-6). Non-CD4 cells were cultured with media only or 100 ng/ml of recombinant IFN-γ. After 72hours, cells were washed and only APCs were irradiated at 3000 rad. After Th17 cells and APC/APC IFN-γ were co-cultured (1∶1) for 16 hrs, the culture supernatant was measured for IL-17 ELISA (right). (b) Splenocytes derived from IFN-γ KO mice were cultured solely or with combined condition with 10 ng/ml of IL-23, 50 ng/ml of IFN-γ, 100 µM 1-MT for 72 hrs. IFN-γ and 1-MT was pretreated. (c) IL-17 mRNA expression of Th17 polarized cells derived from B6 and IDO KO mice splenocyte. (d) Splenocytes derived from IL-1Ra KO mice were cultured solely or with combined condition with 10 ng/ml of IL-23, 50 ng/ml of IFN-γ, 100 µM 1-MT for 72 hrs. IFN-γ and 1-MT was pretreated. ANOVA with post hoc analysis (d,f) was used. Values are presented as the mean ± standard deviation of three independent experiments. *, p

    Journal: PLoS ONE

    Article Title: Interferon Gamma Suppresses Collagen-Induced Arthritis by Regulation of Th17 through the Induction of Indoleamine-2,3-Deoxygenase

    doi: 10.1371/journal.pone.0060900

    Figure Lengend Snippet: Suppression of IL-17 by IFN- γ is associated with IDO. (a) CD4 + T cells isolated from wild type (WT) C57BL/6 (B6) mice and IFN-γ knock out (KO) B6 mice. The cells were cultured in Th17-polarizing conditions (anti CD3 mAb, anti CD28 mAb, anti IFN-γAb, anti IL-4 Ab, TGF-β, IL-6). Non-CD4 cells were cultured with media only or 100 ng/ml of recombinant IFN-γ. After 72hours, cells were washed and only APCs were irradiated at 3000 rad. After Th17 cells and APC/APC IFN-γ were co-cultured (1∶1) for 16 hrs, the culture supernatant was measured for IL-17 ELISA (right). (b) Splenocytes derived from IFN-γ KO mice were cultured solely or with combined condition with 10 ng/ml of IL-23, 50 ng/ml of IFN-γ, 100 µM 1-MT for 72 hrs. IFN-γ and 1-MT was pretreated. (c) IL-17 mRNA expression of Th17 polarized cells derived from B6 and IDO KO mice splenocyte. (d) Splenocytes derived from IL-1Ra KO mice were cultured solely or with combined condition with 10 ng/ml of IL-23, 50 ng/ml of IFN-γ, 100 µM 1-MT for 72 hrs. IFN-γ and 1-MT was pretreated. ANOVA with post hoc analysis (d,f) was used. Values are presented as the mean ± standard deviation of three independent experiments. *, p

    Article Snippet: For intracellular FACS staining, cells were fixed with Cytofix/cytoperm solution (BD) followed by washing with permeablization buffer (BD) and then stained with PE anti mouse IL-17 antibody (ebioscience).

    Techniques: Isolation, Mouse Assay, Knock-Out, Cell Culture, Recombinant, Irradiation, Enzyme-linked Immunosorbent Assay, Derivative Assay, Expressing, Standard Deviation

    The proportion of naïve T cells increased in IFN-γ/IL-17 double knockout mice compared to that of IFN-γ KO mice. Spleen, draining lymph nodes and mesenteric lymph node were removed at 5 th week after 1 st immunization, and prepared for single cell suspension. They were stained with PerCP - anti CD4 ab, APC - anti CD44 ab, and FITC - anti CD62L ab for flowcytometry. (a) Data represents one of three experiments. (b) CD4 + CD44 high CD62L low (memoryCD4 + T cells) T cell population was decreased in CIA (IFN-γIL-17 DKO) group while CD4 + CD44 low CD62L high (naïve CD4 + T cells) population was increased. (c) IL-2, IL-15 and IL-21 were strongly expressed in the joint synovium of CIA (IFN-γ KO). Original magnification, 200x. Mann-Whitney U test used (b). Values are presented as the mean ± standard deviation of four independent experiments. *, p

    Journal: PLoS ONE

    Article Title: Interferon Gamma Suppresses Collagen-Induced Arthritis by Regulation of Th17 through the Induction of Indoleamine-2,3-Deoxygenase

    doi: 10.1371/journal.pone.0060900

    Figure Lengend Snippet: The proportion of naïve T cells increased in IFN-γ/IL-17 double knockout mice compared to that of IFN-γ KO mice. Spleen, draining lymph nodes and mesenteric lymph node were removed at 5 th week after 1 st immunization, and prepared for single cell suspension. They were stained with PerCP - anti CD4 ab, APC - anti CD44 ab, and FITC - anti CD62L ab for flowcytometry. (a) Data represents one of three experiments. (b) CD4 + CD44 high CD62L low (memoryCD4 + T cells) T cell population was decreased in CIA (IFN-γIL-17 DKO) group while CD4 + CD44 low CD62L high (naïve CD4 + T cells) population was increased. (c) IL-2, IL-15 and IL-21 were strongly expressed in the joint synovium of CIA (IFN-γ KO). Original magnification, 200x. Mann-Whitney U test used (b). Values are presented as the mean ± standard deviation of four independent experiments. *, p

    Article Snippet: For intracellular FACS staining, cells were fixed with Cytofix/cytoperm solution (BD) followed by washing with permeablization buffer (BD) and then stained with PE anti mouse IL-17 antibody (ebioscience).

    Techniques: Double Knockout, Mouse Assay, Staining, MANN-WHITNEY, Standard Deviation

    IFN-γ deficient mice demonstrated increased IL-17 production and severe phenotype of collagen-induced arthritis (CIA). Splenocytes from WT B6 or IFN-γ KO B6 mice were cultured with media alone and with 0.5 µg/ml of plate bounded-anti CD3 monoclonal antibodies for 48 hrs (a) to 72 hrs (b) and stained with PE conjugated anti-IL-17 antibody. The proportion of IL-17 producing cells was measured using flow cytometry. (c) Splenocytes from WT B6 or IFN-γ KO B6 mice were cultured in a Th17 polarizing condition. The mRNA expression of IL-17 and the protein level of IL-17 in the supernatant were measured. (d) Sections from the hind paws of WT B6 with or without CIA and IFN-γ KO B6 with or without CIA were obtained at 9 th week after primary CII+CFA immunization. Joint tissue sections were stained with Hematoxylin and Eosin, Toluidine Blue, Safranin O. Infiltration of immune cells in joint synovium between tibia and tarsal bone was observed using microscope and scored according to the referred standard. (e) IL-17 was significantly high in CIA (IFN-γ KO). Joint sections were fixed with paraffin to perform Immunohistochemistry. Original magnification, 400x. (f) Splenocytes from WT B6 with CIA and IFN-γ KO B6 mice were cultured with media alone, 50 µg/ml of CII, 0.5 µg/ml of plate-bounded anti CD3 mAb for 96 hrs, then cell proliferative responses were determined by 3 H-thymidine incorporation assay. Data is represented as the mean counts per minute (CPM). (g) Splenocytes from WT B6 with CIA and IFN-γ KO B6 with CIA were stimulated with 50 µg/ml of CII for 72 hrs. The level of IL-17 in the culture supernatant was measured using ELISA. Mann-Whitney U test (b,c,g) and ANOVA with post hoc analysis (d,f) was used. Values are presented as the mean ± standard deviation of three independent experiments. *, p

    Journal: PLoS ONE

    Article Title: Interferon Gamma Suppresses Collagen-Induced Arthritis by Regulation of Th17 through the Induction of Indoleamine-2,3-Deoxygenase

    doi: 10.1371/journal.pone.0060900

    Figure Lengend Snippet: IFN-γ deficient mice demonstrated increased IL-17 production and severe phenotype of collagen-induced arthritis (CIA). Splenocytes from WT B6 or IFN-γ KO B6 mice were cultured with media alone and with 0.5 µg/ml of plate bounded-anti CD3 monoclonal antibodies for 48 hrs (a) to 72 hrs (b) and stained with PE conjugated anti-IL-17 antibody. The proportion of IL-17 producing cells was measured using flow cytometry. (c) Splenocytes from WT B6 or IFN-γ KO B6 mice were cultured in a Th17 polarizing condition. The mRNA expression of IL-17 and the protein level of IL-17 in the supernatant were measured. (d) Sections from the hind paws of WT B6 with or without CIA and IFN-γ KO B6 with or without CIA were obtained at 9 th week after primary CII+CFA immunization. Joint tissue sections were stained with Hematoxylin and Eosin, Toluidine Blue, Safranin O. Infiltration of immune cells in joint synovium between tibia and tarsal bone was observed using microscope and scored according to the referred standard. (e) IL-17 was significantly high in CIA (IFN-γ KO). Joint sections were fixed with paraffin to perform Immunohistochemistry. Original magnification, 400x. (f) Splenocytes from WT B6 with CIA and IFN-γ KO B6 mice were cultured with media alone, 50 µg/ml of CII, 0.5 µg/ml of plate-bounded anti CD3 mAb for 96 hrs, then cell proliferative responses were determined by 3 H-thymidine incorporation assay. Data is represented as the mean counts per minute (CPM). (g) Splenocytes from WT B6 with CIA and IFN-γ KO B6 with CIA were stimulated with 50 µg/ml of CII for 72 hrs. The level of IL-17 in the culture supernatant was measured using ELISA. Mann-Whitney U test (b,c,g) and ANOVA with post hoc analysis (d,f) was used. Values are presented as the mean ± standard deviation of three independent experiments. *, p

    Article Snippet: For intracellular FACS staining, cells were fixed with Cytofix/cytoperm solution (BD) followed by washing with permeablization buffer (BD) and then stained with PE anti mouse IL-17 antibody (ebioscience).

    Techniques: Mouse Assay, Cell Culture, Staining, Flow Cytometry, Cytometry, Expressing, Microscopy, Immunohistochemistry, Thymidine Incorporation Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Standard Deviation

    In vivo neutralization of interleukin (IL)-17 does not protect against diabetes transferred by diabetic splenocytes. (a) Splenocytes from 15-week-old female diabetic or age- and sex-matched non-diabetic non-obese diabetic (NOD) mice were stimulated in

    Journal: Immunology

    Article Title: Interleukin-17-producing ??+ T cells protect NOD mice from type 1 diabetes through a mechanism involving transforming growth factor-?

    doi: 10.1111/j.1365-2567.2009.03166.x

    Figure Lengend Snippet: In vivo neutralization of interleukin (IL)-17 does not protect against diabetes transferred by diabetic splenocytes. (a) Splenocytes from 15-week-old female diabetic or age- and sex-matched non-diabetic non-obese diabetic (NOD) mice were stimulated in

    Article Snippet: They were then washed, fixed overnight with Fix/Perm buffer (eBioscience), washed with permeabilization buffer (eBioscience), and stained with PE-conjugated anti-IL-17 antibodies (clone FJK-16s; eBioscience) and analysed on a FACScalibur flow cytometer.

    Techniques: In Vivo, Neutralization, Mouse Assay

    Interleukin (IL)-17 expression in non-obese diabetic (NOD) mice is dominated by CD4 − CD8 − γδ + T cells. (a, b) Freshly isolated (a) or IL-23-expanded (b) splenocytes from female NOD mice were double-stained with fluorescein

    Journal: Immunology

    Article Title: Interleukin-17-producing ??+ T cells protect NOD mice from type 1 diabetes through a mechanism involving transforming growth factor-?

    doi: 10.1111/j.1365-2567.2009.03166.x

    Figure Lengend Snippet: Interleukin (IL)-17 expression in non-obese diabetic (NOD) mice is dominated by CD4 − CD8 − γδ + T cells. (a, b) Freshly isolated (a) or IL-23-expanded (b) splenocytes from female NOD mice were double-stained with fluorescein

    Article Snippet: They were then washed, fixed overnight with Fix/Perm buffer (eBioscience), washed with permeabilization buffer (eBioscience), and stained with PE-conjugated anti-IL-17 antibodies (clone FJK-16s; eBioscience) and analysed on a FACScalibur flow cytometer.

    Techniques: Expressing, Mouse Assay, Isolation, Staining

    Interleukin (IL)-23-expanded diabetic splenocytes produced higher levels of IL-17, but induced comparable diabetes in non-obese diabetic–severe combined immunodeficiency (NOD-SCID) mice compared with non-treated diabetic splenocytes. (a, b) Diabetic

    Journal: Immunology

    Article Title: Interleukin-17-producing ??+ T cells protect NOD mice from type 1 diabetes through a mechanism involving transforming growth factor-?

    doi: 10.1111/j.1365-2567.2009.03166.x

    Figure Lengend Snippet: Interleukin (IL)-23-expanded diabetic splenocytes produced higher levels of IL-17, but induced comparable diabetes in non-obese diabetic–severe combined immunodeficiency (NOD-SCID) mice compared with non-treated diabetic splenocytes. (a, b) Diabetic

    Article Snippet: They were then washed, fixed overnight with Fix/Perm buffer (eBioscience), washed with permeabilization buffer (eBioscience), and stained with PE-conjugated anti-IL-17 antibodies (clone FJK-16s; eBioscience) and analysed on a FACScalibur flow cytometer.

    Techniques: Produced, Mouse Assay