mouse il 27  (Thermo Fisher)


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    Name:
    Mouse IL 27 ELISA Standard Recombinant Protein
    Description:
    Mouse IL 27 ELISA Standard Recombinant Protein for ELISA Ctrl
    Catalog Number:
    39-8271-65
    Price:
    None
    Category:
    Proteins Enzymes Peptides
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    Structured Review

    Thermo Fisher mouse il 27
    Mouse IL 27 ELISA Standard Recombinant Protein for ELISA Ctrl
    https://www.bioz.com/result/mouse il 27/product/Thermo Fisher
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    94/100 stars

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    Multiplex Assay:

    Article Title: Rapid activation of tumor-associated macrophages boosts preexisting tumor immunity
    Article Snippet: Protein levels were adjusted after quantification using a bicinchoninic acid protein assay kit (Thermo Fisher) to 10 mg/ml. .. Cytokines and chemokines were determined using a multiplex BioPlex Pro kit (mouse chemokine panel 33-plex and cytokine 23-plex; Biorad) or by ELISA for mouse IL-27 (Invitrogen). .. Readout was performed using BioPlex 200 (Biorad) or Synergy 2 (BioTek) instruments, respectively.

    Bio-Plex Pro Assay:

    Article Title: Rapid activation of tumor-associated macrophages boosts preexisting tumor immunity
    Article Snippet: Protein levels were adjusted after quantification using a bicinchoninic acid protein assay kit (Thermo Fisher) to 10 mg/ml. .. Cytokines and chemokines were determined using a multiplex BioPlex Pro kit (mouse chemokine panel 33-plex and cytokine 23-plex; Biorad) or by ELISA for mouse IL-27 (Invitrogen). .. Readout was performed using BioPlex 200 (Biorad) or Synergy 2 (BioTek) instruments, respectively.

    Enzyme-linked Immunosorbent Assay:

    Article Title: Rapid activation of tumor-associated macrophages boosts preexisting tumor immunity
    Article Snippet: Protein levels were adjusted after quantification using a bicinchoninic acid protein assay kit (Thermo Fisher) to 10 mg/ml. .. Cytokines and chemokines were determined using a multiplex BioPlex Pro kit (mouse chemokine panel 33-plex and cytokine 23-plex; Biorad) or by ELISA for mouse IL-27 (Invitrogen). .. Readout was performed using BioPlex 200 (Biorad) or Synergy 2 (BioTek) instruments, respectively.

    Article Title: Elevated Levels of T-helper 17-associated Cytokines in Diabetes Type I Patients: Indicators for Following the Course of Disease.
    Article Snippet: .. Type 1 diabetes (T1D) is thought to involve chronic inflammation, which is manifested by the activation and expression of different inflammatory mediators. .. Type 1 diabetes (T1D) is thought to involve chronic inflammation, which is manifested by the activation and expression of different inflammatory mediators.

    other:

    Article Title: IRF-8 Controls Melanoma Progression by Regulating the Cross Talk between Cancer and Immune Cells within the Tumor Microenvironment 1IRF-8 Controls Melanoma Progression by Regulating the Cross Talk between Cancer and Immune Cells within the Tumor Microenvironment 1 2
    Article Snippet: Levels of IL-27 were determined by ELISA (eBioscience, San Diego, CA).

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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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    Average 99 stars, based on 1 article reviews
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    93
    Thermo Fisher anti mouse ebi3 antibody
    Over-expression of <t>EBI3</t> blocks IL-12p70 production in CHO cells. A. Biscistronic IL-12p40 and p35 expression were compared before being introduced into a CHO cell by transfection. Two clones (C1 and C2) with comparable levels of IL-12p40 and IL-12p70 as detected by ELISA were selected. B. The biological activity of IL-12 was detected by inducing IFNγ production in C57/black mouse CD4 + T cells. C. The EBI3 expression plasmid was transfected into IL-12p70 expression clone 1 to detect EBI3 expression by Western blotting. Three clones (C1–1, C1–2 and C1–3) with high levels of EBI3 expression plus a non-transfected IL-12p70 expression clone (C1) were used to detect EBI3 expression by Western blotting. The culture supernatants from these 4 clones, plus wild type CHO cells were harvested for use in the IL-12p70 ELISA. D. The culture supernatants were also used to detect IL-35 (EBI3 and p35) heterodimeric protein secretion by immune precipitation Western blotting.
    Anti Mouse Ebi3 Antibody, 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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    93
    Thermo Fisher interleukin 27
    <t>Interleukin‐27</t> ( IL ‐27) induces indoleamine 2,3‐dioxygenase ( IDO ) expression. (a) Neonatal macrophages were treated with IL ‐27 as indicated for 24 hr. RNA was harvested and quantitative PCR performed for IDO gene
    Interleukin 27, 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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    99
    Thermo Fisher mouse monoclonal anti map2
    Characterization of neuronal and astrocytic CLU protein isoforms. ( A: left panel) Schematic of known murine CLU mRNA transcripts provided in the NCBI database. ( A: right panel) Specific CLU gene transcript levels were examined using 25 ng cDNA isolated from DIV 9/16 neurons/astrocytes. Amplicons were visualized on a 4% agarose gel to ensure the appropriate size (left panel) and a comparison of CLU amplicon intensity was visualized (right panel). ( B ) Primary neurons (left panel) or astrocytes (right panel) were prepared as indicated. At DIV 9/16, cytosolic and nuclear fractions were isolated. 30 μg of each fraction was analyzed for CLU protein expression using anti-CLU H-330 and anti-CLU M-18. Fraction purity was analyzed using Gapdh (cytosol), Grp75 (mitochondria), and lamin A (nucleus). Isolation of cell type was confirmed by double labeling with either <t>MAP2</t> (neurons) or GFAP (astrocytes) and CLU H-330, as indicated in the Materials and methods. Cells were imaged at 40X (air; neurons) and 60X (oil; astrocytes) using a customized Olympus IX81/spinning disk confocal inverted microscope and analyzed using the Slidebook Software.
    Mouse Monoclonal Anti Map2, 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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    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

    Over-expression of EBI3 blocks IL-12p70 production in CHO cells. A. Biscistronic IL-12p40 and p35 expression were compared before being introduced into a CHO cell by transfection. Two clones (C1 and C2) with comparable levels of IL-12p40 and IL-12p70 as detected by ELISA were selected. B. The biological activity of IL-12 was detected by inducing IFNγ production in C57/black mouse CD4 + T cells. C. The EBI3 expression plasmid was transfected into IL-12p70 expression clone 1 to detect EBI3 expression by Western blotting. Three clones (C1–1, C1–2 and C1–3) with high levels of EBI3 expression plus a non-transfected IL-12p70 expression clone (C1) were used to detect EBI3 expression by Western blotting. The culture supernatants from these 4 clones, plus wild type CHO cells were harvested for use in the IL-12p70 ELISA. D. The culture supernatants were also used to detect IL-35 (EBI3 and p35) heterodimeric protein secretion by immune precipitation Western blotting.

    Journal: PLoS ONE

    Article Title: Lipopolysaccharide-Induced M2 to M1 Macrophage Transformation for IL-12p70 Production Is Blocked by Candida albicans Mediated Up-Regulation of EBI3 Expression

    doi: 10.1371/journal.pone.0063967

    Figure Lengend Snippet: Over-expression of EBI3 blocks IL-12p70 production in CHO cells. A. Biscistronic IL-12p40 and p35 expression were compared before being introduced into a CHO cell by transfection. Two clones (C1 and C2) with comparable levels of IL-12p40 and IL-12p70 as detected by ELISA were selected. B. The biological activity of IL-12 was detected by inducing IFNγ production in C57/black mouse CD4 + T cells. C. The EBI3 expression plasmid was transfected into IL-12p70 expression clone 1 to detect EBI3 expression by Western blotting. Three clones (C1–1, C1–2 and C1–3) with high levels of EBI3 expression plus a non-transfected IL-12p70 expression clone (C1) were used to detect EBI3 expression by Western blotting. The culture supernatants from these 4 clones, plus wild type CHO cells were harvested for use in the IL-12p70 ELISA. D. The culture supernatants were also used to detect IL-35 (EBI3 and p35) heterodimeric protein secretion by immune precipitation Western blotting.

    Article Snippet: The macrophages were then harvested and washed with cold PBS before staining with anti-mouse EBI3 antibody (eBioscience) or isotype control antibody, followed by PE-Texas red-labelled secondary antibody.

    Techniques: Over Expression, Expressing, Transfection, Clone Assay, Enzyme-linked Immunosorbent Assay, Activity Assay, Plasmid Preparation, Western Blot

    Heat-killed C. albicans enhances EBI3 expression in M2 macrophages with or without LPS stimulation and IL-35 receptors for cytokine consumption. A. M2 macrophages were stimulated with 10 ng/ml LPS or 10 6 cells/ml HKC for 6 hours before lysing the cells for RNA extraction to detect relative levels of EBI3 and IL-12p35 mRNA expression relative to the HPRT mouse housekeeping gene, by RT-qPCR. The cells were stimulated overnight to detect EBI3 and p35 protein expression via Western blotting. B. M2 macrophages were stimulated with increasing amounts of HKC plus 10 ng/ml LPS for 6 hours. The relevant level of EBI3 expression was quantified by RT-qPCR against the HPRT housekeeping gene. EBI3 protein expression was visualised by Western blotting after overnight cell stimulation. The results are representative of two experiments. C. Mouse mIL-12Rβ2 and gp130 mRNA expression in M2 macrophages with or without HKC stimulation. The results are representative of 3 independent experiments. D. Detection of EBI3 on the cell surface of M2 macrophages with and without HKC stimulation.

    Journal: PLoS ONE

    Article Title: Lipopolysaccharide-Induced M2 to M1 Macrophage Transformation for IL-12p70 Production Is Blocked by Candida albicans Mediated Up-Regulation of EBI3 Expression

    doi: 10.1371/journal.pone.0063967

    Figure Lengend Snippet: Heat-killed C. albicans enhances EBI3 expression in M2 macrophages with or without LPS stimulation and IL-35 receptors for cytokine consumption. A. M2 macrophages were stimulated with 10 ng/ml LPS or 10 6 cells/ml HKC for 6 hours before lysing the cells for RNA extraction to detect relative levels of EBI3 and IL-12p35 mRNA expression relative to the HPRT mouse housekeeping gene, by RT-qPCR. The cells were stimulated overnight to detect EBI3 and p35 protein expression via Western blotting. B. M2 macrophages were stimulated with increasing amounts of HKC plus 10 ng/ml LPS for 6 hours. The relevant level of EBI3 expression was quantified by RT-qPCR against the HPRT housekeeping gene. EBI3 protein expression was visualised by Western blotting after overnight cell stimulation. The results are representative of two experiments. C. Mouse mIL-12Rβ2 and gp130 mRNA expression in M2 macrophages with or without HKC stimulation. The results are representative of 3 independent experiments. D. Detection of EBI3 on the cell surface of M2 macrophages with and without HKC stimulation.

    Article Snippet: The macrophages were then harvested and washed with cold PBS before staining with anti-mouse EBI3 antibody (eBioscience) or isotype control antibody, followed by PE-Texas red-labelled secondary antibody.

    Techniques: Expressing, RNA Extraction, Quantitative RT-PCR, Western Blot, Cell Stimulation

    Interleukin‐27 ( IL ‐27) induces indoleamine 2,3‐dioxygenase ( IDO ) expression. (a) Neonatal macrophages were treated with IL ‐27 as indicated for 24 hr. RNA was harvested and quantitative PCR performed for IDO gene

    Journal: Immunology

    Article Title: Elevated interleukin‐27 levels in human neonatal macrophages regulate indoleamine dioxygenase in a STAT‐1 and STAT‐3‐dependent manner

    doi: 10.1111/imm.12625

    Figure Lengend Snippet: Interleukin‐27 ( IL ‐27) induces indoleamine 2,3‐dioxygenase ( IDO ) expression. (a) Neonatal macrophages were treated with IL ‐27 as indicated for 24 hr. RNA was harvested and quantitative PCR performed for IDO gene

    Article Snippet: Interleukin‐27 was measured using the human IL‐27 ELISA kit (eBioscience, San Diego, CA) according to recommended protocol.

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Interleukin‐27 ( IL ‐27) increases indoleamine 2,3‐dioxygenase ( IDO ) activity in neonatal macrophages. (a) Macrophages were treated with IL ‐27 or left untreated ( MED ) as indicated for 72 hr. (b) Macrophages were treated

    Journal: Immunology

    Article Title: Elevated interleukin‐27 levels in human neonatal macrophages regulate indoleamine dioxygenase in a STAT‐1 and STAT‐3‐dependent manner

    doi: 10.1111/imm.12625

    Figure Lengend Snippet: Interleukin‐27 ( IL ‐27) increases indoleamine 2,3‐dioxygenase ( IDO ) activity in neonatal macrophages. (a) Macrophages were treated with IL ‐27 or left untreated ( MED ) as indicated for 72 hr. (b) Macrophages were treated

    Article Snippet: Interleukin‐27 was measured using the human IL‐27 ELISA kit (eBioscience, San Diego, CA) according to recommended protocol.

    Techniques: Activity Assay

    Signal transducer and activator of transcription 1 ( STAT ‐1) and STAT ‐3 are both recruited to the indoleamine 2,3‐dioxygenase ( IDO ) regulatory region. (a) Neonatal macrophages were treated with interleukin‐27 ( IL ‐27)

    Journal: Immunology

    Article Title: Elevated interleukin‐27 levels in human neonatal macrophages regulate indoleamine dioxygenase in a STAT‐1 and STAT‐3‐dependent manner

    doi: 10.1111/imm.12625

    Figure Lengend Snippet: Signal transducer and activator of transcription 1 ( STAT ‐1) and STAT ‐3 are both recruited to the indoleamine 2,3‐dioxygenase ( IDO ) regulatory region. (a) Neonatal macrophages were treated with interleukin‐27 ( IL ‐27)

    Article Snippet: Interleukin‐27 was measured using the human IL‐27 ELISA kit (eBioscience, San Diego, CA) according to recommended protocol.

    Techniques:

    IL‐27 regulates IDO expression

    Journal: Immunology

    Article Title: Elevated interleukin‐27 levels in human neonatal macrophages regulate indoleamine dioxygenase in a STAT‐1 and STAT‐3‐dependent manner

    doi: 10.1111/imm.12625

    Figure Lengend Snippet: IL‐27 regulates IDO expression

    Article Snippet: Interleukin‐27 was measured using the human IL‐27 ELISA kit (eBioscience, San Diego, CA) according to recommended protocol.

    Techniques: Expressing

    Hypothetical model. In response to low or endogenous levels of interleukin‐27 ( IL ‐27), more signal transducer and activator of transcription 3 ( STAT ‐3) is present at the indoleamine 2,3‐dioxygenase ( IDO ) regulatory region.

    Journal: Immunology

    Article Title: Elevated interleukin‐27 levels in human neonatal macrophages regulate indoleamine dioxygenase in a STAT‐1 and STAT‐3‐dependent manner

    doi: 10.1111/imm.12625

    Figure Lengend Snippet: Hypothetical model. In response to low or endogenous levels of interleukin‐27 ( IL ‐27), more signal transducer and activator of transcription 3 ( STAT ‐3) is present at the indoleamine 2,3‐dioxygenase ( IDO ) regulatory region.

    Article Snippet: Interleukin‐27 was measured using the human IL‐27 ELISA kit (eBioscience, San Diego, CA) according to recommended protocol.

    Techniques:

    Elevated interleukin‐27 ( IL ‐27) production by neonatal macrophages. (a) Supernatants from neonatal ( n = 11) and adult ( n = 8) macrophage cultures were collected and IL ‐27 concentration was determined

    Journal: Immunology

    Article Title: Elevated interleukin‐27 levels in human neonatal macrophages regulate indoleamine dioxygenase in a STAT‐1 and STAT‐3‐dependent manner

    doi: 10.1111/imm.12625

    Figure Lengend Snippet: Elevated interleukin‐27 ( IL ‐27) production by neonatal macrophages. (a) Supernatants from neonatal ( n = 11) and adult ( n = 8) macrophage cultures were collected and IL ‐27 concentration was determined

    Article Snippet: Interleukin‐27 was measured using the human IL‐27 ELISA kit (eBioscience, San Diego, CA) according to recommended protocol.

    Techniques: Concentration Assay

    Interleukin‐27 ( IL ‐27) does not induce indoleamine, 2,3‐dioxygenase ( IDO ) activity in murine macrophages. (a) C57 BL /6 mice ( n = 5 or n = 6) were killed at the indicated time in days following birth.

    Journal: Immunology

    Article Title: Elevated interleukin‐27 levels in human neonatal macrophages regulate indoleamine dioxygenase in a STAT‐1 and STAT‐3‐dependent manner

    doi: 10.1111/imm.12625

    Figure Lengend Snippet: Interleukin‐27 ( IL ‐27) does not induce indoleamine, 2,3‐dioxygenase ( IDO ) activity in murine macrophages. (a) C57 BL /6 mice ( n = 5 or n = 6) were killed at the indicated time in days following birth.

    Article Snippet: Interleukin‐27 was measured using the human IL‐27 ELISA kit (eBioscience, San Diego, CA) according to recommended protocol.

    Techniques: Activity Assay, Mouse Assay

    Characterization of neuronal and astrocytic CLU protein isoforms. ( A: left panel) Schematic of known murine CLU mRNA transcripts provided in the NCBI database. ( A: right panel) Specific CLU gene transcript levels were examined using 25 ng cDNA isolated from DIV 9/16 neurons/astrocytes. Amplicons were visualized on a 4% agarose gel to ensure the appropriate size (left panel) and a comparison of CLU amplicon intensity was visualized (right panel). ( B ) Primary neurons (left panel) or astrocytes (right panel) were prepared as indicated. At DIV 9/16, cytosolic and nuclear fractions were isolated. 30 μg of each fraction was analyzed for CLU protein expression using anti-CLU H-330 and anti-CLU M-18. Fraction purity was analyzed using Gapdh (cytosol), Grp75 (mitochondria), and lamin A (nucleus). Isolation of cell type was confirmed by double labeling with either MAP2 (neurons) or GFAP (astrocytes) and CLU H-330, as indicated in the Materials and methods. Cells were imaged at 40X (air; neurons) and 60X (oil; astrocytes) using a customized Olympus IX81/spinning disk confocal inverted microscope and analyzed using the Slidebook Software.

    Journal: eLife

    Article Title: Brain clusterin protein isoforms and mitochondrial localization

    doi: 10.7554/eLife.48255

    Figure Lengend Snippet: Characterization of neuronal and astrocytic CLU protein isoforms. ( A: left panel) Schematic of known murine CLU mRNA transcripts provided in the NCBI database. ( A: right panel) Specific CLU gene transcript levels were examined using 25 ng cDNA isolated from DIV 9/16 neurons/astrocytes. Amplicons were visualized on a 4% agarose gel to ensure the appropriate size (left panel) and a comparison of CLU amplicon intensity was visualized (right panel). ( B ) Primary neurons (left panel) or astrocytes (right panel) were prepared as indicated. At DIV 9/16, cytosolic and nuclear fractions were isolated. 30 μg of each fraction was analyzed for CLU protein expression using anti-CLU H-330 and anti-CLU M-18. Fraction purity was analyzed using Gapdh (cytosol), Grp75 (mitochondria), and lamin A (nucleus). Isolation of cell type was confirmed by double labeling with either MAP2 (neurons) or GFAP (astrocytes) and CLU H-330, as indicated in the Materials and methods. Cells were imaged at 40X (air; neurons) and 60X (oil; astrocytes) using a customized Olympus IX81/spinning disk confocal inverted microscope and analyzed using the Slidebook Software.

    Article Snippet: Sections were incubated overnight at 4°C in PBST supplemented with 1% NGS and mouse monoclonal anti-MAP2 (1:1000, Thermo Fisher Cat. #MA1-25043, Lot #RB2160802), rabbit polyclonal anti-Clusterin H-330 (1:500, Santa Cruz Cat. # sc-8354, Lot # F0316), and rat polyclonal anti-GFAP (1:500, Calbiochem Cat. #345860).

    Techniques: Isolation, Agarose Gel Electrophoresis, Amplification, Expressing, Labeling, Inverted Microscopy, Software