Chronic pancreatitis was induced by repeated acute pancreatitis12 (link). In brief, sex and age matched mice (6-8 weeks old) received 6 hourly intraperitoneal injection of 50μg/kg cerulein (Sigma-Aldrich, St Louis, MO) 3 days/week for a total of 4 weeks. Mice were sacrificed 3 days after the last cerulein injection as described13 (link). For STING agonist treatment, mice were intraperitoneal injected with vehicle or 10mg/kg DMXAA (MedChem Express, New Jersey, USA) daily during the last 5 days of cerulein injection14 (link). For antibody neutralizing experiments, mice were treated with either isotype control or anti-mouse IL-17A (anti-IL-17A, 50μg/mouse/day, 3 times/week; Bio X Cell, NH, USA) antibodies during the last 2 weeks15 (link). To determine STING expression in leukocyte subsets (macrophages and Th17 cells) over time, mice receiving repeated cerulein or saline control (6 hourly injection/day, 3 days/week) were euthanized at week 1, 2, or 3.
Anti il 17a
Manufactured by BioXCell
Sourced in United States
Anti-IL-17A is a laboratory reagent that detects the presence and measures the levels of Interleukin-17A (IL-17A) in biological samples. IL-17A is a pro-inflammatory cytokine that plays a role in various immune and inflammatory processes.
Automatically generated - may contain errors
Lab products found in correlation
Cerulein, by Merck Group (1 mentions)
Dmxaa, by MedChemExpress (1 mentions)
Cerulein, by MedChemExpress (1 mentions)
Anti il 22, by Genentech (1 mentions)
Cd 1 outbred mice, by Charles River Laboratories (1 mentions)
Isotype control clone mopc 21, by BioXCell (1 mentions)
Anti il 17a clone 17f3, by BioXCell (1 mentions)
D luciferin, by Biosynth (1 mentions)
3 protocols using anti il 17a
1
Cerulein-induced Chronic Pancreatitis Mouse Model
2
Isolation and Analysis of Colonic Epithelial Cells
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Primary epithelial cells were isolated from steady state colon of C57BL/6 Rag1−/− mice. Briefly, colons were cut open longitudinally, washed in cold PBS to remove colon content and cut into 1 cm pieces. Epithelial cells were liberated by a 10-min wash with RPMI containing 10% FCS and 5 mM EDTA in a shaking incubator. Supernatant was filtered, cells pelleted at 1,500 r.p.m. and plated at 2 × 105 cells per well in flat bottom 96-well plates in complete media. Recombinant IL-17 and IL-22 (Biolegend) were added for 4 h at the concentration indicated after which cells were harvested and lysed for qPCR.
In some experiments, CD4+TCR-β+CD45.1+ (WT progeny) and CD4+TCR-β+CD45.1− (Tbx21−/− progeny) T cells were sort-purified from the colon of C57BL/6 Rag1−/− recipient mice. Cells were stimulated with IL-23 (10 ng ml−1) for 12 h, the supernatant collected and added at 20% of total culture volume to epithelial cells in combination with blocking antibodies as indicated: anti-IL-17A (10 μg ml−1, clone 17F3 BioXCell), anti-IL-22 (10 μg ml−1, Genentech) or isotype control. Cells were stimulated for 4 h after which cells were harvested and lysed for qPCR.
In some experiments, CD4+TCR-β+CD45.1+ (WT progeny) and CD4+TCR-β+CD45.1− (Tbx21−/− progeny) T cells were sort-purified from the colon of C57BL/6 Rag1−/− recipient mice. Cells were stimulated with IL-23 (10 ng ml−1) for 12 h, the supernatant collected and added at 20% of total culture volume to epithelial cells in combination with blocking antibodies as indicated: anti-IL-17A (10 μg ml−1, clone 17F3 BioXCell), anti-IL-22 (10 μg ml−1, Genentech) or isotype control. Cells were stimulated for 4 h after which cells were harvested and lysed for qPCR.
3
Murine Toxoplasmosis Infection Model
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The luciferase-expressing PRU-Luc-GFP type II of T. gondii strain was provided by J. Boothroyd (Stanford University, Palo Alto, CA). Tissue cysts were obtained from the brains of CD1 outbred mice (Charles River) after 3 mo of infection with 20 cysts by i.p. injection. Experimental mice were infected with 5 cysts of T. gondii PRU-Luc-GFP orally. Weight was monitored every 48 h for the first 20 d and twice a week until day 50. Parasite burden was analyzed by bioluminescence measurements. Mice were imaged every 48 h for 20 d by i.p. injection of 150 mg of luciferin D (Biosynth AG) per kilogram of body weight and using a Xenogen IVIS 100 (Caliper Life Sciences). Data were analyzed with the Living Image Software (Caliper Life Sciences) and is expressed in relative light units. Antibody blockade was performed using anti–IL-17A (clone 17F3; Bio X Cell), isotype control (clone MOPC-21, Bio X Cell), and, in some experiments, anti–IL-17A and anti–IL-17F antibodies (provided by W. Ouyen, Genentech, South San Francisco, CA). For those experiments, mice were injected i.p. with 350 µg antibodies at days −1, 5, and 10 of T. gondii infection.
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