Saline (1.0 mL/kg body weight), MCT oil (1.0 mL/kg body weight) was administered to high-LCT diet (45% fat by energy, 4.73 kcal/g, Cat# D12451) (Research Diets Inc., New Brunswick, NJ, US)-fed, 6-week-old male WT mice and GIP KO mice once daily at 8:00 p.m. by oral gavage. In long-term high-LCT diet tolerance test using a CCK antagonist, drinking water or drinking water supplemented with the CCK antagonist proglumide (0.1 mg/mL, approximately 30 mg/kg/day) (Sigma- Aldrich Japan, Tokyo, Japan) was administered. Body weights were measured once a week. Insulin tolerance test (ITT) and oral glucose tolerance test (OGTT) were performed at weeks 10 and 12 of loading. Fat mass and energy expenditure were measured using computed tomography (CT) and an indirect calorimeter, respectively, at week 14 of loading.
Proglumide
Manufactured by Merck Group
Sourced in United States
Proglumide is a laboratory equipment product manufactured by Merck Group. It is a gastrointestinal agent that functions as a cholecystokinin receptor antagonist.
Automatically generated - may contain errors
Lab products found in correlation
Formic acid, by Thermo Fisher Scientific (3 mentions)
4 nitrobenzoic acid 4 nba, by Merck Group (1 mentions)
Optima grade, by Thermo Fisher Scientific (1 mentions)
Glp 1, by MedChemExpress (1 mentions)
Glp 1, by Merck Group (1 mentions)
Ranitidine, by Merck Group (1 mentions)
2 deoxy d glucose, by Merck Group (1 mentions)
Lorglumide, by Abcam (1 mentions)
Imark microplate absorbance reader, by Bio-Rad (1 mentions)
6 protocols using proglumide
1
High-LCT Diet Tolerance in GIP KO Mice
2
Reagents for Chromatographic Analysis
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The following reagents were purchased: acetonitrile, isopropanol, and methanol from Optima Grade (Fisher Scientific, Waltham, MA, USA); high-purity formic acid (Thermo Scientific, Waltham, MA, USA); proglumide; and 4-nitrobenzoic acid (4-NBA) (Sigma-Aldrich, St. Louis, MI, USA).
3
Pharmacological agents dosages for in-vivo studies
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DON (purity >98%) was obtained from Sigma-Aldrich (St. Louis, MA, USA). DON was dissolved in sterile phosphate buffered saline (PBS) containing 1% dimethyl sulfoxide (DMSO) and dissolved to 0.1, 0.5, 1, 2.5, and 5 mg/kg·bw. GIP (MedChemExpress; Trenton, NJ, USA), PYY, and CCK (Sigma-Aldrich; St. Louis, MO, USA) were respectively diluted in PBS, at a dose of 0.1 mg/kg·bw. GLP-1 (MedChemExpress) was made into a 0.01 mg/kg·bw solution with PBS. The PYY receptor antagonist JNJ-31020028, the CCK receptor antagonist Proglumide (Sigma-Aldrich), and GLP-1 receptor antagonist Exendin(9-39) were respectively dissolved in 100 μL sterile PBS, so that their final doses were 10, 200, or 0.1 mg/kg·bw. GIP receptor inhibition GIP(3-30)NH2 was synthesized by the Syn peptide (Shanghai, China), diluted with normal saline to 50 nm/kg·bw, and injected through the tail vein. According to the instructions of the reagent supplier, previous animal research and thesis data, the dosages of various pharmacological agents were selected [27 (link),31 (link),34 (link),35 (link)].
4
Immunohistochemical Analysis of CCK Receptors
5
Investigating NMDA-Mediated Gastric Secretion
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In the study of NMDA influence on stimulated GAS the following secretagogues were used, being injected i.p., intravenous (i.v.) and intraduodenal (i.d.): carbachol (10 µg/kg, i.p.), histamine (3 mg/kg, i.v.), pentagastrin (0.26 mg/kg, i.p.), cytisine (0.34 mg/kg, i.d., Sopharma, Bulgaria), neutral monocomponent human insulin Acrtapid MC (1.2 u/kg, i.v.), 2-deoxy-D-glucose (100 mg/kg, i.v.) from Sigma Aldrich, USA. The GAS blockers, pentamine (3.2 µg/kg, i.p.), atropine (1 g/kg, i.p.), gastrozepin (3 mg/kg, i.p.), proglumide (10 mg/kg, i.p.), and ranitidine (2 mg/kg, i.p.) were used at doses offered by the manufacturers; AP5 (50 µg/kg, i.p.) (Sigma Aldrich, USA). We used NMDA as an agonist of NMDARs (3 mg/kg, i.p.)27 (link) (Sigma Aldrich, USA). Prior to injections, the drugs were dissolved in 0.9% saline and then heated to 37 °C.
6
Cell Proliferation Assay with Glutamine Antagonists
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Cells were seeded into each well of a 96-well plate (B16-F1 and A375 cells, 2 × 103 cells per well; A431 and HSC-1 cells, 4 × 103 cells per well). After overnight culture at 37 °C, the cells were treated with lorglumide (Abcam), proglumide (Sigma-Aldrich, St. Louis, MO), or loxiglumide (Sigma-Aldrich) for 48 hours. A total of 10 μl of water-soluble tetrazolium salt-8 (Dojindo Molecular Technologies, Kumamoto, Japan) was added to each well of a 96-well plate, and the cells were incubated for another 3 hours at 37 °C. Optical density was measured using an iMark Microplate Absorbance Reader (Bio-Rad Laboratory) at 450 nm.
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