Female mice were randomly divided into the control group, DNFB group, I3C group, CCFM1029 group, and AHR antagonist CH223191 (Sigma-Aldrich) group (n = 8). One week before model establishment, the mice were gavaged B. longum CCFM1029 suspension (diluting to 109 CFU/0.2 mL with sterile saline) once a day in the CH223191 and CCFM1029 groups until the end of the experiment, and 10 μM CH223191 (20 μL) was painted on the ear and skin for 30 min before B. longum CCFM1029 treatment.18 (link) The mice were gavaged 0.2 mL sterile saline in the control and DNFB groups, and oral administration of 0.2 mL I3C solution (10 μg/mL, Sigma-Aldrich) in the I3C group.
Ch223191
Manufactured by Merck Group
Sourced in United States, Germany, Italy
CH223191 is a laboratory equipment product. It is used for scientific research and analysis purposes. The core function of this product is to facilitate the measurement and analysis of various samples and materials in a laboratory setting. However, a detailed description of its specific features and capabilities cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Dimethyl sulfoxide (dmso), by Merck Group (14 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (5 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Vancomycin, by Merck Group (4 mentions)
Ampicillin, by Merck Group (4 mentions)
Hepes, by Thermo Fisher Scientific (4 mentions)
Indole, by Merck Group (3 mentions)
Kynurenine, by Merck Group (3 mentions)
Rpmi 1640, by Thermo Fisher Scientific (3 mentions)
L glutamine, by Thermo Fisher Scientific (3 mentions)
Dimethyl sulfoxide (dmso), by Fujifilm (3 mentions)
Penicillin, by Thermo Fisher Scientific (3 mentions)
Streptomycin, by Thermo Fisher Scientific (3 mentions)
2 3 7 8 tetrachlorodibenzo p dioxin tcdd, by Cambridge Isotopes (2 mentions)
Benzo a pyrene, by Merck Group (2 mentions)
6 formylindolo 3 2 b carbazole ficz, by Enzo Life Sciences (2 mentions)
Actinomycin d, by Merck Group (2 mentions)
Neomycin sulfate, by Merck Group (2 mentions)
Metronidazole, by Merck Group (2 mentions)
Ingenuity pathway analysis (ipa), by Merck Group (2 mentions)
98 protocols using ch223191
1
Modulation of Allergic Dermatitis by Bifidobacterium longum
2
Oral Gavage Probiotic Modulation
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All mice were treated by oral gavage daily thorough out the course of the experiments. Mice received indole by gavage (20 mg/kg/dose or ~200 μL of 17 mM indole; Sigma Aldrich, St. Louis, MO) dissolved in sterile water warmed to 55 °C. All probiotics (Bifidobacterium bifidum, Bifidobacterium lactis, Lactobacillus plantarum, Lactobacillus reuteri, and Lactobacillus rhamnosus) were obtained from DuPont™ Danisco® FloraFIT® probiotics as lyophilized powder at a known CFU/gram concentration. Each probiotic was weight and aliquoted into 1 × 108 CFU aliquots. Probiotics or heat-killed probiotics were resuspended in PBS and mice were gavaged with 5 × 108 CFU/dose. Probiotic cocktails were heat-killed by incubating at 100 °C for 15 min, bacterial viability was then confirmed by plating. No viable organisms were recovered from the heat-killed probiotics. Mice received CH-223191 by gavage (10 mg/kg/dose or ~200 µL of 3 mM CH-223191; Sigma Aldrich) dissolved in sterile corn oil. Alcohol-fed and pair-fed mice were gavage daily with vehicle (PBS). A separated cohort of control mice was given corn-oil or water daily to ensure no effects were seen with the different vehicles. No changes in host defense or bacterial burden were seen in control mice given corn-oil or water.
3
Investigating UV-B Damage Pathways
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This experiment was performed to obtain further insights into the contribution of two independent vitamin D-dependent signaling pathways that are mediated by the classical vitamin D receptor (VDR) and the aryl hydrocarbon receptor (AhR) for the cellular response to UV-B-induced damage. We applied an AhR-Antagonist CH223191 (Sigma) at a final concentration of 10−7 M, achieved by diluting an ethanol stock solution (10−4 M) at 1:1000 in medium and a partial VDR-Inhibitor (25-Hydroxyvitamin D3 [25(OH)2D3, Sigma]) at the same final concentration of 10−7 M. Previous studies [29 (link),30 (link),31 (link)] have confirmed the effective blocking of the AhR and VDR receptors by CH223191 and 25(OH)2D3, respectively, at this concentration. We used all 16 possible combinations of treatment factors UVB, 1,25(OH)2D3, CH223191, and 25(OH)2D3 as illustrated in Table 1 . As the control (standard) we used an extra dish treated with Triton-X Solution (Merck Millipore, Burlington, VT, USA) diluted down to 1% in DMEM containing 1% BSA and incubated for 15′. LDH activity was measured with a Cytotoxicity Detection Kit (Roche, Mannheim, Germany). The LDH activity measured in the free medium of treated cells was directly proportional to the level of toxicity undergone by the cells under their respective treatments [32 (link)].
4
Embryonic Xenobiotic Response Assay
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In stimulation experiments, 2 dpf embryos were treated with different compounds in E3 medium for the durations indicated in the respective figures. In experiments using the AhR inhibitor CH-223191 (Sigma-Aldrich), embryos were pre-exposed to 10 mM CH-223191 in E3 for 2 h prior to the experiment and the inhibitor was present during the entire duration of the experiment.
5
Characterization of RAW 264.7 Macrophages
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RAW 264.7 cells were purchased from ATCC (Manassas, MA, USA). Cells were authenticated and tested for mycoplasma contamination using the CellCheck 19 Plus test (ATCC). Dulbecco’s Modified Eagle Medium (DMEM), penicillin/streptomycin, and LPS (from Salmonella minnesota) were purchased from Invitrogen (Carlsbad, CA, USA). Fetal bovine serum (FBS) was purchased from Atlanta Biologicals (Flowery Branch, GA, USA). All FFA chemicals, AICAR, and the AhR inhibitor CH-223191 were purchased from Millipore Sigma (St. Louis, MO, USA). I3A sodium salt was purchased from Cayman chemicals (Ann Arbor, MI, USA). All other chemicals were purchased from VWR (Radnor, PA, USA) or Millipore Sigma unless otherwise specified.
6
Cell Culture Conditions for AhR Studies
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Karpas 299, Jurkat, and Peer cells were propagated in Roswell Park Memorial Institute (RPMI) 1640 Medium, Hepa-1c1c7 cells were cultured in Minimum Essential Medium (MEM) Alpha, and stable BpRc1 cells were cultured in Dulbecco's Modified Eagle Medium (DMEM) and 2 μg/mL puromycin (InvivoGen). For CK2 inhibitor experiments, cells were treated with 5 μM CX-4945 (Selleckchem) for 1 h prior to AhR ligand exposure. AhR antagonist experiments were conducted by pretreating cells with 10 μM CH223191 (MilliporeSigma) for 2 h prior to AhR ligand exposure. Culture and reagent details are described in SI Appendix .
7
Antioxidant and AhR Activity Assay
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Chemicals 2-Hydroxychrysene (purity > 99%) and 6-hydroxychrysene (purity > 99%) were purchased from Toronto Research Chemicals, Ontario, Canada. The AhR antagonist, CH-223191, (purity ≥ 98%), α-tocopherol (purity ≥ 96%), N-acetyl-L-cysteine (purity ≥ 99%), 2,6-di-tert-butyl-4-methylphenol (purity ≥ 99%), 2thiobarbituric acid (purity ≥ 98%), 1,1,3,3-tetramethoxypropane (purity ≥ 99%), and 2′,7′-dichlorofluorescin diacetate (purity ≥ 97%) were purchased from Millipore-Sigma. 2-Thiobarbituric acid was dissolved in 100 mM NaOH and 1,1,3,3tetramethoxypropane was dissolved in 1.15% KCl. The rest of the compounds were dissolved in dimethyl sulfoxide (DMSO). Exposure solutions were made fresh daily by dilution of stock solutions to 0.1% DMSO in deionized (DI) water.
8
Pharmacological Characterization of Signaling Pathways
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9
Exposure of Cells to Diesel Exhaust
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Human recombinant IL-1β was purchased from Merck (Prague, Czech Republic) and reconstituted according to the supplier’s instructions. Stock solutions were stored at −20 °C. Dimethyl sulfoxide (DMSO) and CH-223191 were obtained from Merck. 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) was purchased from Cambridge Isotope Laboratories (Andover, MA, USA) and dissolved in DMSO. Standard reference mixture of diesel exhaust particles (DEP) produced by a combination of direct injection four-cycle diesel engines SRM 1650b [26 ] was purchased from the National Institute of Standards and Technology (Gaithersberg, MD, USA). It was extracted and fractionated based on different molecule polarity to separate different classes of polycyclic aromatic compounds, as previously described [27 (link),28 (link)]. Both crude SRM1650b extract (CE) and its polar (F3) fraction were used for further analyses, at non-cytotoxic concentration (0.1 mg SRM equivalents/mL), which was selected based on preliminary experiments. All other chemicals were obtained from Merck, if not stated otherwise.
10
Investigating Oxidative Stress Responses
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FICZ (Enzo Life Sciences, Farmingdale, NY) was dissolved in dimethyl sulfoxide (DMSO; Sigma-Aldrich, St. Louis, MO) and added to Dulbecco's modified Eagle's medium (DMEM; Sigma-Aldrich) at final concentrations of 1, 10, 100, and 1000 nM. Benzo[a]pyrene (BaP; Sigma-Aldrich) and N-acetyl-L-cysteine (NAC; Sigma-Aldrich) were dissolved in DMSO and added to the culture medium at final concentrations of 1 and 5 μM, respectively. An oxidation-sensitive dye, carboxy-H2DCFDA (Thermo Fisher Scientific, Waltham, MA), was dissolved in DMSO at a concentration of 10 mM and further diluted in HBSS (Fujifilm Wako Pure Chemical Corporation, Osaka, Japan) at a final concentration of 25 μM. CH223191 (Merck, Darmstadt, Germany), an AHR antagonist, was dissolved in DMSO and added to culture medium at a final concentration of 10 μM. The antibodies used were rabbit anti-human β-actin antibody (Cell Signaling Technology, Danvers, MA), rabbit anti-human AHR antibody (H-211) (Santa Cruz Biotechnology, Dallas, TX), rabbit anti-human NF-κB p65 (Abcam, Cambridge, United Kingdom), and horseradish-peroxidase-conjugated anti-rabbit secondary antibody (Cell Signaling Technology).
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