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rat anti mouse il 36α antibody  (R&D Systems)


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    R&D Systems rat anti mouse il 36α antibody
    A) Coomassie blue stained gel demonstrating the purity of <t>IL-36α</t> preparation. Ten µg IL-36α was loaded on the lane. B) Western immunoblotting of IL-36α protein preparation detects a band around 18 KDa, the predicted molecular weight of mouse IL-36α. C) Cytospin preparations demonstrating neutrophil influx in the BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. D) Total cell counts from BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. E) Differential cell count percentages and F) Differential cell count numbers in the BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. *Indicates significant differences ( P<0.05 ) compared to PBS treated mice. Data represent mean±SEM from 4–5 mice/group. G) Flow cytometry on cells recovered from BAL fluid from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. A majority of cells from the IL-36α instilled lungs were CD11c − CD11b + Ly6G + neutrophils. Depicted flow cytometry plots are representative of 4–5 mice/group. H) Hematoxylin & Eosin stained lung sections isolated from mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Depicted sections are representative of 4–5 mice/group.
    Rat Anti Mouse Il 36α Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rat anti mouse il 36α antibody/product/R&D Systems
    Average 93 stars, based on 2 article reviews
    rat anti mouse il 36α antibody - by Bioz Stars, 2026-03
    93/100 stars

    Images

    1) Product Images from "IL-36α Exerts Pro-Inflammatory Effects in the Lungs of Mice"

    Article Title: IL-36α Exerts Pro-Inflammatory Effects in the Lungs of Mice

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0045784

    A) Coomassie blue stained gel demonstrating the purity of IL-36α preparation. Ten µg IL-36α was loaded on the lane. B) Western immunoblotting of IL-36α protein preparation detects a band around 18 KDa, the predicted molecular weight of mouse IL-36α. C) Cytospin preparations demonstrating neutrophil influx in the BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. D) Total cell counts from BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. E) Differential cell count percentages and F) Differential cell count numbers in the BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. *Indicates significant differences ( P<0.05 ) compared to PBS treated mice. Data represent mean±SEM from 4–5 mice/group. G) Flow cytometry on cells recovered from BAL fluid from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. A majority of cells from the IL-36α instilled lungs were CD11c − CD11b + Ly6G + neutrophils. Depicted flow cytometry plots are representative of 4–5 mice/group. H) Hematoxylin & Eosin stained lung sections isolated from mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Depicted sections are representative of 4–5 mice/group.
    Figure Legend Snippet: A) Coomassie blue stained gel demonstrating the purity of IL-36α preparation. Ten µg IL-36α was loaded on the lane. B) Western immunoblotting of IL-36α protein preparation detects a band around 18 KDa, the predicted molecular weight of mouse IL-36α. C) Cytospin preparations demonstrating neutrophil influx in the BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. D) Total cell counts from BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. E) Differential cell count percentages and F) Differential cell count numbers in the BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. *Indicates significant differences ( P<0.05 ) compared to PBS treated mice. Data represent mean±SEM from 4–5 mice/group. G) Flow cytometry on cells recovered from BAL fluid from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. A majority of cells from the IL-36α instilled lungs were CD11c − CD11b + Ly6G + neutrophils. Depicted flow cytometry plots are representative of 4–5 mice/group. H) Hematoxylin & Eosin stained lung sections isolated from mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Depicted sections are representative of 4–5 mice/group.

    Techniques Used: Staining, Western Blot, Molecular Weight, Cell Counting, Flow Cytometry, Isolation

    A–H) Transcript expression of early response cytokines (TNFα, IL-1α, IL-1β, IL-36γ), the classical IL-1 receptor IL-1R1, the novel IL-1 cytokine cluster receptor IL-36R and the neutrophil specific chemokines CXCL1 and CXCL2 in the lungs of mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Transcript expression was evaluated by SYBR-Green based quantitative real-time PCR. I–K) Protein expression of TNFα, IL-1α, IL-1β and CXCL1 in the BAL fluid recovered from mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Protein expression was quantified by multiplexed cytometric bead arrays. *Indicates significant differences ( P<0.05 ) compared to PBS treated mice. Data represent mean±SEM from 4–5 mice/group.
    Figure Legend Snippet: A–H) Transcript expression of early response cytokines (TNFα, IL-1α, IL-1β, IL-36γ), the classical IL-1 receptor IL-1R1, the novel IL-1 cytokine cluster receptor IL-36R and the neutrophil specific chemokines CXCL1 and CXCL2 in the lungs of mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Transcript expression was evaluated by SYBR-Green based quantitative real-time PCR. I–K) Protein expression of TNFα, IL-1α, IL-1β and CXCL1 in the BAL fluid recovered from mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Protein expression was quantified by multiplexed cytometric bead arrays. *Indicates significant differences ( P<0.05 ) compared to PBS treated mice. Data represent mean±SEM from 4–5 mice/group.

    Techniques Used: Expressing, SYBR Green Assay, Real-time Polymerase Chain Reaction

    A) Total lung resistance and B) lung compliance were not significantly different in the lungs of IL-36α challenged mice compared to PBS controls. Airway responses in mice were measured using invasive plethysmography 24 h following i.t instillation of IL-36α or PBS. Data presented are percentage changes from baseline (0mg/mL) measurements. Data represent mean±SEM from 5–7 mice/group.
    Figure Legend Snippet: A) Total lung resistance and B) lung compliance were not significantly different in the lungs of IL-36α challenged mice compared to PBS controls. Airway responses in mice were measured using invasive plethysmography 24 h following i.t instillation of IL-36α or PBS. Data presented are percentage changes from baseline (0mg/mL) measurements. Data represent mean±SEM from 5–7 mice/group.

    Techniques Used:

    A) Total cell counts from BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. B) Differential cell count percentages and C) Differential cell count numbers in the BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. *Indicates significant differences ( P<0.05 ) compared to PBS treated mice. Data represent mean±SEM from 3–4 mice/group. D) Flow cytometry on cells recovered from BAL fluid from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. A majority of cells from the IL-36α instilled lungs were CD11c − CD11b + Ly6G + neutrophils. Depicted flow cytometry plots are representative of 3–4 mice/group. H) Hematoxylin & Eosin stained lung sections isolated from mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Depicted sections are representative of 3–4 mice/group.
    Figure Legend Snippet: A) Total cell counts from BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. B) Differential cell count percentages and C) Differential cell count numbers in the BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. *Indicates significant differences ( P<0.05 ) compared to PBS treated mice. Data represent mean±SEM from 3–4 mice/group. D) Flow cytometry on cells recovered from BAL fluid from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. A majority of cells from the IL-36α instilled lungs were CD11c − CD11b + Ly6G + neutrophils. Depicted flow cytometry plots are representative of 3–4 mice/group. H) Hematoxylin & Eosin stained lung sections isolated from mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Depicted sections are representative of 3–4 mice/group.

    Techniques Used: Cell Counting, Flow Cytometry, Staining, Isolation

    A–F) Transcript expression of early response cytokines (TNFα and IL-36γ), the classical IL-1 receptor IL-1R1, the novel IL-1 cytokine cluster receptor IL-36R and the neutrophil specific chemokines CXCL1 and CXCL2 in the lungs of IL-1αβ −/− mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Transcript expression was evaluated by SYBR-Green based quantitative real-time PCR. G–H) Protein expression of TNFα and CXCL1 in the BAL fluid recovered from IL-1αβ −/− mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Protein expression was quantified by multiplexed cytometric bead arrays. *Indicates significant differences ( P<0.05 ) compared to PBS treated mice. Data represent mean±SEM from 3–4 mice/group.
    Figure Legend Snippet: A–F) Transcript expression of early response cytokines (TNFα and IL-36γ), the classical IL-1 receptor IL-1R1, the novel IL-1 cytokine cluster receptor IL-36R and the neutrophil specific chemokines CXCL1 and CXCL2 in the lungs of IL-1αβ −/− mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Transcript expression was evaluated by SYBR-Green based quantitative real-time PCR. G–H) Protein expression of TNFα and CXCL1 in the BAL fluid recovered from IL-1αβ −/− mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Protein expression was quantified by multiplexed cytometric bead arrays. *Indicates significant differences ( P<0.05 ) compared to PBS treated mice. Data represent mean±SEM from 3–4 mice/group.

    Techniques Used: Expressing, SYBR Green Assay, Real-time Polymerase Chain Reaction

    A–H) Transcript expression of early response cytokines (TNFα, IL-1α, IL-1β, IL-36γ), the classical IL-1 receptor IL-1R1, the novel IL-1 cytokine cluster receptor IL-36R as well as the neutrophil specific chemokines CXCL1 and CXCL2 in splenic CD11c + cells 2 h following incubation with increasing concentrations of IL-36α. Transcript expression was evaluated by SYBR-Green based quantitative real-time PCR. *Indicates significant differences ( P<0.05 ) compared to 0 µg/mL group. Data represent mean±SD of quadruplicate samples from one of two representative experiments. I) Diff-quik stained cells from cytospun BAL cells from the lungs of naïve mice demonstrates that the majority of lung resident immune cells are alveolar macrophages. J) PCR on cDNA from naïve mouse alveolar macrophages demonstrating the constitutive mRNA expression of an endogenous control (β-actin), IL-1R1, IL-36R and IL-1RAcP. Image of a DNA electrophoresis gel has been color-inverted for clarity. bp – base pairs.
    Figure Legend Snippet: A–H) Transcript expression of early response cytokines (TNFα, IL-1α, IL-1β, IL-36γ), the classical IL-1 receptor IL-1R1, the novel IL-1 cytokine cluster receptor IL-36R as well as the neutrophil specific chemokines CXCL1 and CXCL2 in splenic CD11c + cells 2 h following incubation with increasing concentrations of IL-36α. Transcript expression was evaluated by SYBR-Green based quantitative real-time PCR. *Indicates significant differences ( P<0.05 ) compared to 0 µg/mL group. Data represent mean±SD of quadruplicate samples from one of two representative experiments. I) Diff-quik stained cells from cytospun BAL cells from the lungs of naïve mice demonstrates that the majority of lung resident immune cells are alveolar macrophages. J) PCR on cDNA from naïve mouse alveolar macrophages demonstrating the constitutive mRNA expression of an endogenous control (β-actin), IL-1R1, IL-36R and IL-1RAcP. Image of a DNA electrophoresis gel has been color-inverted for clarity. bp – base pairs.

    Techniques Used: Expressing, Incubation, SYBR Green Assay, Real-time Polymerase Chain Reaction, Diff-Quik, Staining, Control, Nucleic Acid Electrophoresis

    A–C) Transcript expression of the co-stimulatory molecules CD80, CD86 and CD40 in splenic CD11c + cells 2 h following incubation with increasing concentrations of IL-36α. Transcript expression was evaluated by SYBR-Green based quantitative real-time PCR. *Indicates significant differences ( P<0.05 ) compared to 0 µg/mL group. Data represent mean±SD from quadruplicate samples from one of two representative experiments. D) Flow cytometric evaluation of splenic CD11c + cells 24 h following incubation with increasing concentrations of IL-36α for 2 h. E) Cell surface expression of co-stimulatory molecules in splenic CD11c + cells 24 h following incubation with increasing concentrations of IL-36α for 2 h. MFI – mean fluorescence intensity. *Indicates significant differences ( P<0.05 ) compared to 0.1 µg/mL group. Data represent mean±SD from triplicate samples from one of two representative experiments.
    Figure Legend Snippet: A–C) Transcript expression of the co-stimulatory molecules CD80, CD86 and CD40 in splenic CD11c + cells 2 h following incubation with increasing concentrations of IL-36α. Transcript expression was evaluated by SYBR-Green based quantitative real-time PCR. *Indicates significant differences ( P<0.05 ) compared to 0 µg/mL group. Data represent mean±SD from quadruplicate samples from one of two representative experiments. D) Flow cytometric evaluation of splenic CD11c + cells 24 h following incubation with increasing concentrations of IL-36α for 2 h. E) Cell surface expression of co-stimulatory molecules in splenic CD11c + cells 24 h following incubation with increasing concentrations of IL-36α for 2 h. MFI – mean fluorescence intensity. *Indicates significant differences ( P<0.05 ) compared to 0.1 µg/mL group. Data represent mean±SD from triplicate samples from one of two representative experiments.

    Techniques Used: Expressing, Incubation, SYBR Green Assay, Real-time Polymerase Chain Reaction, Fluorescence

    A) Flow cytometric evaluation of CD4 + T cell proliferation responses induced by IL-36α stimulated splenic CD11c + cells. Splenic CD11c + cells were incubated with increasing doses of IL-36α for 2 h. Following incubation, media containing IL-36α was removed and CFSE-labeled CD4 + T cells were co-cultured with IL-36α stimulated CD11c + cells. CFSE dilution was used to evaluate T cell proliferation responses 96 h following co-culture. CD4 + T cell proliferation was proportional to the concentration of IL-36α used for stimulating CD11c + cells used in the co-culture. Flow cytometry plot presented is representative of quadruplicate samples in one out of two independent experiments. B) Flow cytometric evaluation of antigen-specific CD4 + T cell proliferation responses induced by IL-36α stimulated splenic CD11c + cells. Splenic CD11c + cells were incubated with increasing doses of IL-36α and 100 ng/mL OVA 323-339 for 2 h. Following incubation, media containing IL-36α and the OVA peptide was removed and CFSE-labeled CD4 + T cells from OTII TCR transgenic mice were co-cultured with IL-36α stimulated, OVA peptide pulsed CD11c + cells. CFSE dilution in the culture was used to evaluate T cell proliferation responses 96 h following co-culture. CD4 + T cell proliferation was proportional to the concentration of IL-36α used for stimulating CD11c + cells used in the co-culture. Flow cytometry plot presented is representative of quadruplicate samples in one out of two independent experiments.
    Figure Legend Snippet: A) Flow cytometric evaluation of CD4 + T cell proliferation responses induced by IL-36α stimulated splenic CD11c + cells. Splenic CD11c + cells were incubated with increasing doses of IL-36α for 2 h. Following incubation, media containing IL-36α was removed and CFSE-labeled CD4 + T cells were co-cultured with IL-36α stimulated CD11c + cells. CFSE dilution was used to evaluate T cell proliferation responses 96 h following co-culture. CD4 + T cell proliferation was proportional to the concentration of IL-36α used for stimulating CD11c + cells used in the co-culture. Flow cytometry plot presented is representative of quadruplicate samples in one out of two independent experiments. B) Flow cytometric evaluation of antigen-specific CD4 + T cell proliferation responses induced by IL-36α stimulated splenic CD11c + cells. Splenic CD11c + cells were incubated with increasing doses of IL-36α and 100 ng/mL OVA 323-339 for 2 h. Following incubation, media containing IL-36α and the OVA peptide was removed and CFSE-labeled CD4 + T cells from OTII TCR transgenic mice were co-cultured with IL-36α stimulated, OVA peptide pulsed CD11c + cells. CFSE dilution in the culture was used to evaluate T cell proliferation responses 96 h following co-culture. CD4 + T cell proliferation was proportional to the concentration of IL-36α used for stimulating CD11c + cells used in the co-culture. Flow cytometry plot presented is representative of quadruplicate samples in one out of two independent experiments.

    Techniques Used: Incubation, Labeling, Cell Culture, Co-Culture Assay, Concentration Assay, Flow Cytometry, Transgenic Assay

    A) Cells from a mouse macrophage NF-κB reporter cell line (RAW-ELAM cells) were stimulated with increasing concentrations of IL-36α. Green fluorescent protein (GFP) expression, indicative of NF-κB activation, was increased in a dose-dependent manner upon incubation with IL-36α. Flow cytometry plot presented is representative of triplicate samples in one out of two independent experiments.
    Figure Legend Snippet: A) Cells from a mouse macrophage NF-κB reporter cell line (RAW-ELAM cells) were stimulated with increasing concentrations of IL-36α. Green fluorescent protein (GFP) expression, indicative of NF-κB activation, was increased in a dose-dependent manner upon incubation with IL-36α. Flow cytometry plot presented is representative of triplicate samples in one out of two independent experiments.

    Techniques Used: Expressing, Activation Assay, Incubation, Flow Cytometry



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    rat anti mouse il 36α antibody  (R&D Systems)
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    R&D Systems rat anti mouse il 36α antibody
    A) Coomassie blue stained gel demonstrating the purity of <t>IL-36α</t> preparation. Ten µg IL-36α was loaded on the lane. B) Western immunoblotting of IL-36α protein preparation detects a band around 18 KDa, the predicted molecular weight of mouse IL-36α. C) Cytospin preparations demonstrating neutrophil influx in the BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. D) Total cell counts from BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. E) Differential cell count percentages and F) Differential cell count numbers in the BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. *Indicates significant differences ( P<0.05 ) compared to PBS treated mice. Data represent mean±SEM from 4–5 mice/group. G) Flow cytometry on cells recovered from BAL fluid from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. A majority of cells from the IL-36α instilled lungs were CD11c − CD11b + Ly6G + neutrophils. Depicted flow cytometry plots are representative of 4–5 mice/group. H) Hematoxylin & Eosin stained lung sections isolated from mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Depicted sections are representative of 4–5 mice/group.
    Rat Anti Mouse Il 36α Antibody, supplied by R&D Systems, 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/rat anti mouse il 36α antibody/product/R&D Systems
    Average 93 stars, based on 1 article reviews
    rat anti mouse il 36α antibody - by Bioz Stars, 2026-03
    93/100 stars
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    A) Coomassie blue stained gel demonstrating the purity of IL-36α preparation. Ten µg IL-36α was loaded on the lane. B) Western immunoblotting of IL-36α protein preparation detects a band around 18 KDa, the predicted molecular weight of mouse IL-36α. C) Cytospin preparations demonstrating neutrophil influx in the BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. D) Total cell counts from BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. E) Differential cell count percentages and F) Differential cell count numbers in the BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. *Indicates significant differences ( P<0.05 ) compared to PBS treated mice. Data represent mean±SEM from 4–5 mice/group. G) Flow cytometry on cells recovered from BAL fluid from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. A majority of cells from the IL-36α instilled lungs were CD11c − CD11b + Ly6G + neutrophils. Depicted flow cytometry plots are representative of 4–5 mice/group. H) Hematoxylin & Eosin stained lung sections isolated from mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Depicted sections are representative of 4–5 mice/group.

    Journal: PLoS ONE

    Article Title: IL-36α Exerts Pro-Inflammatory Effects in the Lungs of Mice

    doi: 10.1371/journal.pone.0045784

    Figure Lengend Snippet: A) Coomassie blue stained gel demonstrating the purity of IL-36α preparation. Ten µg IL-36α was loaded on the lane. B) Western immunoblotting of IL-36α protein preparation detects a band around 18 KDa, the predicted molecular weight of mouse IL-36α. C) Cytospin preparations demonstrating neutrophil influx in the BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. D) Total cell counts from BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. E) Differential cell count percentages and F) Differential cell count numbers in the BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. *Indicates significant differences ( P<0.05 ) compared to PBS treated mice. Data represent mean±SEM from 4–5 mice/group. G) Flow cytometry on cells recovered from BAL fluid from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. A majority of cells from the IL-36α instilled lungs were CD11c − CD11b + Ly6G + neutrophils. Depicted flow cytometry plots are representative of 4–5 mice/group. H) Hematoxylin & Eosin stained lung sections isolated from mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Depicted sections are representative of 4–5 mice/group.

    Article Snippet: Proteins were transferred to polyvinylidene fluoride membranes, probed with rat anti-mouse IL-36α antibody (R&D Systems, Catalog No. MAB2297) and detected with a horseradish peroxidase conjugated goat anti-rat secondary antibody (Abcam) and ECL Plus chemiluminescent detection system (Amersham).

    Techniques: Staining, Western Blot, Molecular Weight, Cell Counting, Flow Cytometry, Isolation

    A–H) Transcript expression of early response cytokines (TNFα, IL-1α, IL-1β, IL-36γ), the classical IL-1 receptor IL-1R1, the novel IL-1 cytokine cluster receptor IL-36R and the neutrophil specific chemokines CXCL1 and CXCL2 in the lungs of mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Transcript expression was evaluated by SYBR-Green based quantitative real-time PCR. I–K) Protein expression of TNFα, IL-1α, IL-1β and CXCL1 in the BAL fluid recovered from mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Protein expression was quantified by multiplexed cytometric bead arrays. *Indicates significant differences ( P<0.05 ) compared to PBS treated mice. Data represent mean±SEM from 4–5 mice/group.

    Journal: PLoS ONE

    Article Title: IL-36α Exerts Pro-Inflammatory Effects in the Lungs of Mice

    doi: 10.1371/journal.pone.0045784

    Figure Lengend Snippet: A–H) Transcript expression of early response cytokines (TNFα, IL-1α, IL-1β, IL-36γ), the classical IL-1 receptor IL-1R1, the novel IL-1 cytokine cluster receptor IL-36R and the neutrophil specific chemokines CXCL1 and CXCL2 in the lungs of mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Transcript expression was evaluated by SYBR-Green based quantitative real-time PCR. I–K) Protein expression of TNFα, IL-1α, IL-1β and CXCL1 in the BAL fluid recovered from mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Protein expression was quantified by multiplexed cytometric bead arrays. *Indicates significant differences ( P<0.05 ) compared to PBS treated mice. Data represent mean±SEM from 4–5 mice/group.

    Article Snippet: Proteins were transferred to polyvinylidene fluoride membranes, probed with rat anti-mouse IL-36α antibody (R&D Systems, Catalog No. MAB2297) and detected with a horseradish peroxidase conjugated goat anti-rat secondary antibody (Abcam) and ECL Plus chemiluminescent detection system (Amersham).

    Techniques: Expressing, SYBR Green Assay, Real-time Polymerase Chain Reaction

    A) Total lung resistance and B) lung compliance were not significantly different in the lungs of IL-36α challenged mice compared to PBS controls. Airway responses in mice were measured using invasive plethysmography 24 h following i.t instillation of IL-36α or PBS. Data presented are percentage changes from baseline (0mg/mL) measurements. Data represent mean±SEM from 5–7 mice/group.

    Journal: PLoS ONE

    Article Title: IL-36α Exerts Pro-Inflammatory Effects in the Lungs of Mice

    doi: 10.1371/journal.pone.0045784

    Figure Lengend Snippet: A) Total lung resistance and B) lung compliance were not significantly different in the lungs of IL-36α challenged mice compared to PBS controls. Airway responses in mice were measured using invasive plethysmography 24 h following i.t instillation of IL-36α or PBS. Data presented are percentage changes from baseline (0mg/mL) measurements. Data represent mean±SEM from 5–7 mice/group.

    Article Snippet: Proteins were transferred to polyvinylidene fluoride membranes, probed with rat anti-mouse IL-36α antibody (R&D Systems, Catalog No. MAB2297) and detected with a horseradish peroxidase conjugated goat anti-rat secondary antibody (Abcam) and ECL Plus chemiluminescent detection system (Amersham).

    Techniques:

    A) Total cell counts from BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. B) Differential cell count percentages and C) Differential cell count numbers in the BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. *Indicates significant differences ( P<0.05 ) compared to PBS treated mice. Data represent mean±SEM from 3–4 mice/group. D) Flow cytometry on cells recovered from BAL fluid from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. A majority of cells from the IL-36α instilled lungs were CD11c − CD11b + Ly6G + neutrophils. Depicted flow cytometry plots are representative of 3–4 mice/group. H) Hematoxylin & Eosin stained lung sections isolated from mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Depicted sections are representative of 3–4 mice/group.

    Journal: PLoS ONE

    Article Title: IL-36α Exerts Pro-Inflammatory Effects in the Lungs of Mice

    doi: 10.1371/journal.pone.0045784

    Figure Lengend Snippet: A) Total cell counts from BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. B) Differential cell count percentages and C) Differential cell count numbers in the BAL fluid recovered from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. *Indicates significant differences ( P<0.05 ) compared to PBS treated mice. Data represent mean±SEM from 3–4 mice/group. D) Flow cytometry on cells recovered from BAL fluid from mice 24 h following a single i.t. instillation of PBS or 10 µg IL-36α. A majority of cells from the IL-36α instilled lungs were CD11c − CD11b + Ly6G + neutrophils. Depicted flow cytometry plots are representative of 3–4 mice/group. H) Hematoxylin & Eosin stained lung sections isolated from mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Depicted sections are representative of 3–4 mice/group.

    Article Snippet: Proteins were transferred to polyvinylidene fluoride membranes, probed with rat anti-mouse IL-36α antibody (R&D Systems, Catalog No. MAB2297) and detected with a horseradish peroxidase conjugated goat anti-rat secondary antibody (Abcam) and ECL Plus chemiluminescent detection system (Amersham).

    Techniques: Cell Counting, Flow Cytometry, Staining, Isolation

    A–F) Transcript expression of early response cytokines (TNFα and IL-36γ), the classical IL-1 receptor IL-1R1, the novel IL-1 cytokine cluster receptor IL-36R and the neutrophil specific chemokines CXCL1 and CXCL2 in the lungs of IL-1αβ −/− mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Transcript expression was evaluated by SYBR-Green based quantitative real-time PCR. G–H) Protein expression of TNFα and CXCL1 in the BAL fluid recovered from IL-1αβ −/− mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Protein expression was quantified by multiplexed cytometric bead arrays. *Indicates significant differences ( P<0.05 ) compared to PBS treated mice. Data represent mean±SEM from 3–4 mice/group.

    Journal: PLoS ONE

    Article Title: IL-36α Exerts Pro-Inflammatory Effects in the Lungs of Mice

    doi: 10.1371/journal.pone.0045784

    Figure Lengend Snippet: A–F) Transcript expression of early response cytokines (TNFα and IL-36γ), the classical IL-1 receptor IL-1R1, the novel IL-1 cytokine cluster receptor IL-36R and the neutrophil specific chemokines CXCL1 and CXCL2 in the lungs of IL-1αβ −/− mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Transcript expression was evaluated by SYBR-Green based quantitative real-time PCR. G–H) Protein expression of TNFα and CXCL1 in the BAL fluid recovered from IL-1αβ −/− mice 24 h following a single i.t instillation of PBS or 10 µg IL-36α. Protein expression was quantified by multiplexed cytometric bead arrays. *Indicates significant differences ( P<0.05 ) compared to PBS treated mice. Data represent mean±SEM from 3–4 mice/group.

    Article Snippet: Proteins were transferred to polyvinylidene fluoride membranes, probed with rat anti-mouse IL-36α antibody (R&D Systems, Catalog No. MAB2297) and detected with a horseradish peroxidase conjugated goat anti-rat secondary antibody (Abcam) and ECL Plus chemiluminescent detection system (Amersham).

    Techniques: Expressing, SYBR Green Assay, Real-time Polymerase Chain Reaction

    A–H) Transcript expression of early response cytokines (TNFα, IL-1α, IL-1β, IL-36γ), the classical IL-1 receptor IL-1R1, the novel IL-1 cytokine cluster receptor IL-36R as well as the neutrophil specific chemokines CXCL1 and CXCL2 in splenic CD11c + cells 2 h following incubation with increasing concentrations of IL-36α. Transcript expression was evaluated by SYBR-Green based quantitative real-time PCR. *Indicates significant differences ( P<0.05 ) compared to 0 µg/mL group. Data represent mean±SD of quadruplicate samples from one of two representative experiments. I) Diff-quik stained cells from cytospun BAL cells from the lungs of naïve mice demonstrates that the majority of lung resident immune cells are alveolar macrophages. J) PCR on cDNA from naïve mouse alveolar macrophages demonstrating the constitutive mRNA expression of an endogenous control (β-actin), IL-1R1, IL-36R and IL-1RAcP. Image of a DNA electrophoresis gel has been color-inverted for clarity. bp – base pairs.

    Journal: PLoS ONE

    Article Title: IL-36α Exerts Pro-Inflammatory Effects in the Lungs of Mice

    doi: 10.1371/journal.pone.0045784

    Figure Lengend Snippet: A–H) Transcript expression of early response cytokines (TNFα, IL-1α, IL-1β, IL-36γ), the classical IL-1 receptor IL-1R1, the novel IL-1 cytokine cluster receptor IL-36R as well as the neutrophil specific chemokines CXCL1 and CXCL2 in splenic CD11c + cells 2 h following incubation with increasing concentrations of IL-36α. Transcript expression was evaluated by SYBR-Green based quantitative real-time PCR. *Indicates significant differences ( P<0.05 ) compared to 0 µg/mL group. Data represent mean±SD of quadruplicate samples from one of two representative experiments. I) Diff-quik stained cells from cytospun BAL cells from the lungs of naïve mice demonstrates that the majority of lung resident immune cells are alveolar macrophages. J) PCR on cDNA from naïve mouse alveolar macrophages demonstrating the constitutive mRNA expression of an endogenous control (β-actin), IL-1R1, IL-36R and IL-1RAcP. Image of a DNA electrophoresis gel has been color-inverted for clarity. bp – base pairs.

    Article Snippet: Proteins were transferred to polyvinylidene fluoride membranes, probed with rat anti-mouse IL-36α antibody (R&D Systems, Catalog No. MAB2297) and detected with a horseradish peroxidase conjugated goat anti-rat secondary antibody (Abcam) and ECL Plus chemiluminescent detection system (Amersham).

    Techniques: Expressing, Incubation, SYBR Green Assay, Real-time Polymerase Chain Reaction, Diff-Quik, Staining, Control, Nucleic Acid Electrophoresis

    A–C) Transcript expression of the co-stimulatory molecules CD80, CD86 and CD40 in splenic CD11c + cells 2 h following incubation with increasing concentrations of IL-36α. Transcript expression was evaluated by SYBR-Green based quantitative real-time PCR. *Indicates significant differences ( P<0.05 ) compared to 0 µg/mL group. Data represent mean±SD from quadruplicate samples from one of two representative experiments. D) Flow cytometric evaluation of splenic CD11c + cells 24 h following incubation with increasing concentrations of IL-36α for 2 h. E) Cell surface expression of co-stimulatory molecules in splenic CD11c + cells 24 h following incubation with increasing concentrations of IL-36α for 2 h. MFI – mean fluorescence intensity. *Indicates significant differences ( P<0.05 ) compared to 0.1 µg/mL group. Data represent mean±SD from triplicate samples from one of two representative experiments.

    Journal: PLoS ONE

    Article Title: IL-36α Exerts Pro-Inflammatory Effects in the Lungs of Mice

    doi: 10.1371/journal.pone.0045784

    Figure Lengend Snippet: A–C) Transcript expression of the co-stimulatory molecules CD80, CD86 and CD40 in splenic CD11c + cells 2 h following incubation with increasing concentrations of IL-36α. Transcript expression was evaluated by SYBR-Green based quantitative real-time PCR. *Indicates significant differences ( P<0.05 ) compared to 0 µg/mL group. Data represent mean±SD from quadruplicate samples from one of two representative experiments. D) Flow cytometric evaluation of splenic CD11c + cells 24 h following incubation with increasing concentrations of IL-36α for 2 h. E) Cell surface expression of co-stimulatory molecules in splenic CD11c + cells 24 h following incubation with increasing concentrations of IL-36α for 2 h. MFI – mean fluorescence intensity. *Indicates significant differences ( P<0.05 ) compared to 0.1 µg/mL group. Data represent mean±SD from triplicate samples from one of two representative experiments.

    Article Snippet: Proteins were transferred to polyvinylidene fluoride membranes, probed with rat anti-mouse IL-36α antibody (R&D Systems, Catalog No. MAB2297) and detected with a horseradish peroxidase conjugated goat anti-rat secondary antibody (Abcam) and ECL Plus chemiluminescent detection system (Amersham).

    Techniques: Expressing, Incubation, SYBR Green Assay, Real-time Polymerase Chain Reaction, Fluorescence

    A) Flow cytometric evaluation of CD4 + T cell proliferation responses induced by IL-36α stimulated splenic CD11c + cells. Splenic CD11c + cells were incubated with increasing doses of IL-36α for 2 h. Following incubation, media containing IL-36α was removed and CFSE-labeled CD4 + T cells were co-cultured with IL-36α stimulated CD11c + cells. CFSE dilution was used to evaluate T cell proliferation responses 96 h following co-culture. CD4 + T cell proliferation was proportional to the concentration of IL-36α used for stimulating CD11c + cells used in the co-culture. Flow cytometry plot presented is representative of quadruplicate samples in one out of two independent experiments. B) Flow cytometric evaluation of antigen-specific CD4 + T cell proliferation responses induced by IL-36α stimulated splenic CD11c + cells. Splenic CD11c + cells were incubated with increasing doses of IL-36α and 100 ng/mL OVA 323-339 for 2 h. Following incubation, media containing IL-36α and the OVA peptide was removed and CFSE-labeled CD4 + T cells from OTII TCR transgenic mice were co-cultured with IL-36α stimulated, OVA peptide pulsed CD11c + cells. CFSE dilution in the culture was used to evaluate T cell proliferation responses 96 h following co-culture. CD4 + T cell proliferation was proportional to the concentration of IL-36α used for stimulating CD11c + cells used in the co-culture. Flow cytometry plot presented is representative of quadruplicate samples in one out of two independent experiments.

    Journal: PLoS ONE

    Article Title: IL-36α Exerts Pro-Inflammatory Effects in the Lungs of Mice

    doi: 10.1371/journal.pone.0045784

    Figure Lengend Snippet: A) Flow cytometric evaluation of CD4 + T cell proliferation responses induced by IL-36α stimulated splenic CD11c + cells. Splenic CD11c + cells were incubated with increasing doses of IL-36α for 2 h. Following incubation, media containing IL-36α was removed and CFSE-labeled CD4 + T cells were co-cultured with IL-36α stimulated CD11c + cells. CFSE dilution was used to evaluate T cell proliferation responses 96 h following co-culture. CD4 + T cell proliferation was proportional to the concentration of IL-36α used for stimulating CD11c + cells used in the co-culture. Flow cytometry plot presented is representative of quadruplicate samples in one out of two independent experiments. B) Flow cytometric evaluation of antigen-specific CD4 + T cell proliferation responses induced by IL-36α stimulated splenic CD11c + cells. Splenic CD11c + cells were incubated with increasing doses of IL-36α and 100 ng/mL OVA 323-339 for 2 h. Following incubation, media containing IL-36α and the OVA peptide was removed and CFSE-labeled CD4 + T cells from OTII TCR transgenic mice were co-cultured with IL-36α stimulated, OVA peptide pulsed CD11c + cells. CFSE dilution in the culture was used to evaluate T cell proliferation responses 96 h following co-culture. CD4 + T cell proliferation was proportional to the concentration of IL-36α used for stimulating CD11c + cells used in the co-culture. Flow cytometry plot presented is representative of quadruplicate samples in one out of two independent experiments.

    Article Snippet: Proteins were transferred to polyvinylidene fluoride membranes, probed with rat anti-mouse IL-36α antibody (R&D Systems, Catalog No. MAB2297) and detected with a horseradish peroxidase conjugated goat anti-rat secondary antibody (Abcam) and ECL Plus chemiluminescent detection system (Amersham).

    Techniques: Incubation, Labeling, Cell Culture, Co-Culture Assay, Concentration Assay, Flow Cytometry, Transgenic Assay

    A) Cells from a mouse macrophage NF-κB reporter cell line (RAW-ELAM cells) were stimulated with increasing concentrations of IL-36α. Green fluorescent protein (GFP) expression, indicative of NF-κB activation, was increased in a dose-dependent manner upon incubation with IL-36α. Flow cytometry plot presented is representative of triplicate samples in one out of two independent experiments.

    Journal: PLoS ONE

    Article Title: IL-36α Exerts Pro-Inflammatory Effects in the Lungs of Mice

    doi: 10.1371/journal.pone.0045784

    Figure Lengend Snippet: A) Cells from a mouse macrophage NF-κB reporter cell line (RAW-ELAM cells) were stimulated with increasing concentrations of IL-36α. Green fluorescent protein (GFP) expression, indicative of NF-κB activation, was increased in a dose-dependent manner upon incubation with IL-36α. Flow cytometry plot presented is representative of triplicate samples in one out of two independent experiments.

    Article Snippet: Proteins were transferred to polyvinylidene fluoride membranes, probed with rat anti-mouse IL-36α antibody (R&D Systems, Catalog No. MAB2297) and detected with a horseradish peroxidase conjugated goat anti-rat secondary antibody (Abcam) and ECL Plus chemiluminescent detection system (Amersham).

    Techniques: Expressing, Activation Assay, Incubation, Flow Cytometry