rabbit anti human tlr4 polyclonal antibody  (Bioss)

Journal: Frontiers in Nutrition

Article Title: Galactooligosaccharide Treatment Alleviates DSS-Induced Colonic Inflammation in Caco-2 Cell Model

doi: 10.3389/fnut.2022.862974

Figure Lengend Snippet: Inhibition of DSS induced-NF-kB signaling pathway by Bimuno GOS in Caco-2 cells. Cells were untreated, or treated with 2% DSS alone for 6 h, or treated with 2% DSS for 2 h and then with Bimuno GOS (100 or 200 μg/ml) for a further 4 h, or treated with 100 or 200 μg/ml Bimuno GOS alone for 4 h. Protein expression was analyzed by Western blot. The relative expression levels of TLR4, MyD88, Tollip, and IRAK-M were normalized to α-tubulin, whereas the phosphorylated IKK-α, IKK-β, IkB-α, and p65 were normalized to their corresponding unphosphorylated forms. (A) TLR4, MyD88, P-IKK-α, P-IKK-β, P-IkB-α, P-p65 (densitometric values). (B) Tollip and IRAK-M (densitometric values). (C) Representative Western blot of the analyzed proteins. Values represent means ± SD of three independent experiments, carried out in triplicate. Means without a common letter significantly differ, P < 0.05.

Article Snippet: Membranes were incubated with the following primary antibodies: rabbit polyclonal anti-human TLR4, MyD88, IKKα, IKKβ, phospho(P)-IKKα/β, IkBα, P-IkBα, NF-kB p65, P-p65, IRAK-M, Tollip, from Cell Signaling Technology (Danvers, MA), mouse monoclonal α-tubulin.

Techniques: Inhibition, Expressing, Western Blot

Journal: Frontiers in Nutrition

Article Title: Galactooligosaccharide Treatment Alleviates DSS-Induced Colonic Inflammation in Caco-2 Cell Model

doi: 10.3389/fnut.2022.862974

Figure Lengend Snippet: Inhibition of DSS induced-P-p65 translocation by Bimuno GOS in Caco-2 cells. Cells were untreated (C), or treated with 2% DSS for 6 h, or treated with Bimuno GOS (100 or 200 μg/ml) for 4 h, or treated with 2% DSS for 2 h and then with Bimuno GOS (100 or 200 μg/ml) for further 4 h. Cell nuclei were stained with DAPI, while P-p65 was labeled with rabbit polyclonal anti-P-p65 antibody, followed by TRITC-conjugated secondary antibody. Each figure is representative of three independent assays (63 × magnification). Bars represent 10 μm.

Article Snippet: Membranes were incubated with the following primary antibodies: rabbit polyclonal anti-human TLR4, MyD88, IKKα, IKKβ, phospho(P)-IKKα/β, IkBα, P-IkBα, NF-kB p65, P-p65, IRAK-M, Tollip, from Cell Signaling Technology (Danvers, MA), mouse monoclonal α-tubulin.

Techniques: Inhibition, Translocation Assay, Staining, Labeling

Journal: Scientific Reports

Article Title: Hypercapnia Alters Expression of Immune Response, Nucleosome Assembly and Lipid Metabolism Genes in Differentiated Human Bronchial Epithelial Cells

doi: 10.1038/s41598-018-32008-x

Figure Lengend Snippet: Hypercapnia alters expression of genes involved in innate immunity and host defense. ALI-differentiated NHBE cells were exposed to normocapnia (NC) or hypercapnia (HC) for 24 h prior to analysis. ( a ) Cluster A genes altered by hypercapnia and their associated GO biological processes. ( b ) Heatmap and hierarchical clustering of gene expression profiles in normocapnia and hypercapnia. ( c ) CXCL1, CXCL14, CCL28, IL6R , and TLR4 mRNA expression levels were assessed by qPCR and expression in hypercapnia was expressed as fold change relative to normocapnia. Results shown are means ± SE; n = 3. ( d ) Representative fluorescence micrographs of NHBE cells double-stained for TLR4 (red) and the cilia marker acetylated tubulin (green), and counter stained with Hoechst (blue). Non immune control (NIC) cells stained without primary antibodies. ( e ) Immunoblotting of whole cell lysates for TLR4. Histogram shows densitometry of TLR4 normalized to β-actin (loading control). Results shown are means ± SE; n = 3.

Article Snippet: Membranes were probed with polyclonal rabbit anti-human TLR4 (H-80) antibody followed by HRP-conjugated anti-rabbit secondary antibody (Pierce).

Techniques: Expressing, Fluorescence, Staining, Marker, Western Blot

Journal: Scientific Reports

Article Title: Hypercapnia Alters Expression of Immune Response, Nucleosome Assembly and Lipid Metabolism Genes in Differentiated Human Bronchial Epithelial Cells

doi: 10.1038/s41598-018-32008-x

Figure Lengend Snippet: Hypercapnia alters expression of genes involved in innate immunity and host defense. ALI-differentiated NHBE cells were exposed to normocapnia (NC) or hypercapnia (HC) for 24 h prior to analysis. ( a ) Cluster A genes altered by hypercapnia and their associated GO biological processes. ( b ) Heatmap and hierarchical clustering of gene expression profiles in normocapnia and hypercapnia. ( c ) CXCL1, CXCL14, CCL28, IL6R , and TLR4 mRNA expression levels were assessed by qPCR and expression in hypercapnia was expressed as fold change relative to normocapnia. Results shown are means ± SE; n = 3. ( d ) Representative fluorescence micrographs of NHBE cells double-stained for TLR4 (red) and the cilia marker acetylated tubulin (green), and counter stained with Hoechst (blue). Non immune control (NIC) cells stained without primary antibodies. ( e ) Immunoblotting of whole cell lysates for TLR4. Histogram shows densitometry of TLR4 normalized to β-actin (loading control). Results shown are means ± SE; n = 3.

Article Snippet: Cells were blocked in PBS containing 2% BSA and 0.1% triton X-100 then double-stained with 1:200 polyclonal rabbit anti-human TLR4 antibody (H-80, Santa Cruz Biotechnology) followed by 1:200 Alexa Fluor 555-conjugated goat-anti-rabbit IgG (red) (Invitrogen), and 1:500 monoclonal mouse anti-human acetylated tubulin antibody (Clone 6–11B-1, Sigma) followed by 1:200 Alexa fluor 488-conjugated goat anti-mouse IgG (green) (Invitrogen).

Techniques: Expressing, Gene Expression, Fluorescence, Staining, Marker, Control, Western Blot