Saline:Article Title: CD30 knockout attenuates experimental colitis by reducing inflammatory cytokine production
Article Snippet: Equal amounts of protein (30 μg) were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membranes (Millipore, Burlington, MA, USA) using a Trans-Blot system (Bio-Rad, Hercules, CA, USA). .. Membranes were blocked with 5% non-fat milk (BD Biosciences) in Tris-buffered saline with 0.1% Tween-20 (TBST) for 1 h at room temperature and incubated overnight at 4 °C with following primary antibodies (1:1000 dilution): anti-CD30 (Cat. No. AF852; R&D Systems), anti-T-bet (Cat. No. ab91109; Abcam, Cambridge, MA, USA), anti-IFN-γ (Cat. No. ab9657; Abcam), anti-STAT4 (Cat. No. 2653; Cell Signaling Technology, Danvers, MA, USA), anti-phospho-STAT4 (Tyr693) (Cat. No. 4134; Cell Signaling Technology), anti-NF-κB p65 (Cat. No. 8242; Cell Signaling Technology), anti-phospho-NF-κB p65 (Ser536) (Cat. No. 3033; Cell Signaling Technology), anti-IκBα (Cat. No. 4814; Cell Signaling Technology), anti-phospho-IκBα (Ser32) (Cat. No. 2859; Cell Signaling Technology), and anti-β-actin (Cat. No. 4970; Cell Signaling Technology). .. After washing with TBST, membranes were incubated with horseradish peroxidase-conjugated secondary antibodies (Cat. No. 7074 for anti-rabbit IgG, Cat. No. 7076 for anti-mouse IgG; Cell Signaling Technology, 1:5000 dilution) for 1 h at room temperature.
Article Title: CD30 knockout attenuates experimental colitis by reducing inflammatory cytokine production.
Article Snippet: Equal amounts of protein (30 μg) were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membranes (Millipore, Burlington, MA, USA) using a Trans-Blot system (Bio-Rad, Hercules, CA, USA). .. Membranes were blocked with 5% non-fat milk (BD Biosciences) in Tris-buffered saline with 0.1% Tween-20 (TBST) for 1 hour at room temperature and incubated AR TIC LE IN PR ES S overnight at 4°C with following primary antibodies (1:1000 dilution): anti-CD30 (Cat. No. AF852; R&D Systems), anti-T-bet (Cat. No. ab91109; Abcam, Cambridge, MA, USA), anti-IFN-γ (Cat. No. ab9657; Abcam), anti-STAT4 (Cat. No. 2653; Cell Signaling Technology, Danvers, MA, USA), anti-phospho-STAT4 (Tyr693) (Cat. No. 4134; Cell Signaling Technology), anti-NF-κB p65 (Cat. No. 8242; Cell Signaling Technology), anti-phospho-NF-κB p65 (Ser536) (Cat. No. 3033; Cell Signaling Technology), anti-IκBα (Cat. No. 4814; Cell Signaling Technology), anti-phospho-IκBα (Ser32) (Cat. No. 2859; Cell Signaling Technology), and anti-β-actin (Cat. No. 4970; Cell Signaling Technology). .. After washing with TBST, membranes were incubated with horseradish peroxidase-conjugated secondary antibodies (Cat. No. 7074 for anti-rabbit IgG, Cat. No. 7076 for anti-mouse IgG; Cell Signaling Technology, 1:5000 dilution) for 1 hour at room temperature.
Incubation:Article Title: CD30 knockout attenuates experimental colitis by reducing inflammatory cytokine production
Article Snippet: Equal amounts of protein (30 μg) were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membranes (Millipore, Burlington, MA, USA) using a Trans-Blot system (Bio-Rad, Hercules, CA, USA). .. Membranes were blocked with 5% non-fat milk (BD Biosciences) in Tris-buffered saline with 0.1% Tween-20 (TBST) for 1 h at room temperature and incubated overnight at 4 °C with following primary antibodies (1:1000 dilution): anti-CD30 (Cat. No. AF852; R&D Systems), anti-T-bet (Cat. No. ab91109; Abcam, Cambridge, MA, USA), anti-IFN-γ (Cat. No. ab9657; Abcam), anti-STAT4 (Cat. No. 2653; Cell Signaling Technology, Danvers, MA, USA), anti-phospho-STAT4 (Tyr693) (Cat. No. 4134; Cell Signaling Technology), anti-NF-κB p65 (Cat. No. 8242; Cell Signaling Technology), anti-phospho-NF-κB p65 (Ser536) (Cat. No. 3033; Cell Signaling Technology), anti-IκBα (Cat. No. 4814; Cell Signaling Technology), anti-phospho-IκBα (Ser32) (Cat. No. 2859; Cell Signaling Technology), and anti-β-actin (Cat. No. 4970; Cell Signaling Technology). .. After washing with TBST, membranes were incubated with horseradish peroxidase-conjugated secondary antibodies (Cat. No. 7074 for anti-rabbit IgG, Cat. No. 7076 for anti-mouse IgG; Cell Signaling Technology, 1:5000 dilution) for 1 h at room temperature.
Article Title: CD30 knockout attenuates experimental colitis by reducing inflammatory cytokine production.
Article Snippet: Equal amounts of protein (30 μg) were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membranes (Millipore, Burlington, MA, USA) using a Trans-Blot system (Bio-Rad, Hercules, CA, USA). .. Membranes were blocked with 5% non-fat milk (BD Biosciences) in Tris-buffered saline with 0.1% Tween-20 (TBST) for 1 hour at room temperature and incubated AR TIC LE IN PR ES S overnight at 4°C with following primary antibodies (1:1000 dilution): anti-CD30 (Cat. No. AF852; R&D Systems), anti-T-bet (Cat. No. ab91109; Abcam, Cambridge, MA, USA), anti-IFN-γ (Cat. No. ab9657; Abcam), anti-STAT4 (Cat. No. 2653; Cell Signaling Technology, Danvers, MA, USA), anti-phospho-STAT4 (Tyr693) (Cat. No. 4134; Cell Signaling Technology), anti-NF-κB p65 (Cat. No. 8242; Cell Signaling Technology), anti-phospho-NF-κB p65 (Ser536) (Cat. No. 3033; Cell Signaling Technology), anti-IκBα (Cat. No. 4814; Cell Signaling Technology), anti-phospho-IκBα (Ser32) (Cat. No. 2859; Cell Signaling Technology), and anti-β-actin (Cat. No. 4970; Cell Signaling Technology). .. After washing with TBST, membranes were incubated with horseradish peroxidase-conjugated secondary antibodies (Cat. No. 7074 for anti-rabbit IgG, Cat. No. 7076 for anti-mouse IgG; Cell Signaling Technology, 1:5000 dilution) for 1 hour at room temperature.
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