Mice were fasted overnight (14 h). Before starting the experiment, mice were acclimatized to single cages without bedding for at least 1 h. Then, carrier solutions, WIN 55,212-2 (1 mg·kg−1) or loperamide (5 mg·kg−1) were injected i.p. After 20 min, a plastic bead (2 mm in diameter) was inserted 2 cm into the colon under mild anaesthesia using IsoFlo/Isoflurane (Abbott). Then, time until expulsion of the plastic bead was recorded.
Isoflo isoflurane
Manufactured by Abbott
Sourced in United Kingdom, United States
IsoFlo/Isoflurane is a general anesthetic that is administered through inhalation. It is used to induce and maintain general anesthesia in surgical procedures. The product is designed to provide a controlled and consistent delivery of the anesthetic gas to patients.
Automatically generated - may contain errors
Lab products found in correlation
Direct red 80, by Merck Group (2 mentions)
Phosphate buffered saline solution, by Harvard Bioscience (2 mentions)
Ponceau s, by Merck Group (2 mentions)
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Doxorubicin hydrochloride, by Merck Group (2 mentions)
Dc protein assay kit, by Bio-Rad (2 mentions)
Sodium chloride (nacl), by Avantor (1 mentions)
5 5 dithiobis 2 nitrobenzoic acid, by Merck Group (1 mentions)
7 protocols using isoflo isoflurane
1
Colonic Bead Expulsion Assay
2
Mice Husbandry and Experimental Procedures
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Mice were maintained on a regular light-dark cycle (14 h light, 10 h dark) with a room temperature of 23 ± 1°C and kept ad libitum on a standard laboratory chow diet (4.5% w/w fat, Ssniff Spezialdiaeten, Soest, Germany). Animals used for experiments were 12–20 weeks of age. If not mentioned otherwise, male mice were used. MGL-KO mice were generated as described previously (Taschler et al., 2011 (link)). CB1-deficient (CB1-KO) mice were obtained from A. Zimmer (University of Bonn, Bonn, Germany) and generated as described (Zimmer et al., 1999 (link)). Animals were anaesthetized with IsoFlo/Isoflurane (Abbott, Animal Health, Queenborough, Kent, UK) and killed by cervical dislocation. All studies involving animals are reported in accordance with the ARRIVE guidelines for reporting experiments involving animals (Kilkenny et al., 2010 (link); McGrath et al., 2010 (link)). Experimental procedures were approved by the Ethics committee of the University of Graz, the Austrian Federal Ministry of Science and Research (protocol number BMWF 66007/7-ll/3b/2013) and were conducted in accordance with the council of Europe Convention (ETS 123). Procedures were performed as humanely as possible to minimize suffering.
3
Vitamin A-Deficient Mice Metabolism Study
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Mice were maintained on a regular light–dark cycle (12 h light, 12 h dark) and kept ad libitum on a standard laboratory chow diet (Ssniff Spezialdiaeten, Soest, Germany, Vitamin A ~ 15,000 IU/kg) or on a Vitamin A-deficient diet (Ssniff, Vitamin A < 120 IU/kg). ATGL-ko and HSL-ko mice were generated by targeted homologous recombination as described previously [15,16] . Animals were 8–12 weeks of age. Female mice were used for all studies. Non-fasted animals were anesthetized with IsoFlo/Isoflurane (Abbott, Animal Health, Queenborough, Kent, UK) and euthanized by cervical dislocation. The study was approved by the ethics committee of the University of Graz and is in accordance with the Council of Europe Convention (ETS 123).
4
Vitamin A-Deficient Diet Modulates Lipolysis
5
Genetically-modified Hsl knockout mice
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Hsl knockout mice were generated by targeted homologous recombination as described previously (3 (link)) and backcrossed at least 10 times to a C57Bl/6J genetic background. Mice were kept on a standard laboratory chow diet containing 4.5% (w/w) fat (sniff Spezialdiäten GmbH, Soest, Germany). For all studies, male mice with an age range of 10–14 weeks were used. Animals had ad libitum access to food and water and were housed with a light and dark period of 10 and 14 h, respectively, at a room temperature of 22°C ± 1°C. For refeeding conditions, mice were fasted for 12 h and had then free access to food for 2 h. Body mass composition was assessed in nonanesthetized mice using the time-domain NMR minispec (Live Mice Analyzer system, Model LF90II; Bruker Optik, Germany). Animals were anesthetized with ISOflo®/isoflurane (Abbott, Abbott Park, IL) and sacrificed by cervical dislocation. All experiments were approved by the Austrian Federal Ministry for Science and the local ethics committee (BMWF-66.007/0017-II/3b/2013).
6
Doxorubicin-Induced Oxidative Stress Evaluation
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Doxorubicin hydrochloride (≥ 98% purity, DOX), Ponceau S, direct red 80, 5,5-dithiobis(2-nitrobenzoic acid), adenosine triphosphate (ATP), reduced glutathione (GSH), glutathione reductase, oxidized glutathione (GSSG) disodium salt, bovine serum albumin, and the all other chemicals used were purchased from Sigma-Aldrich (St. Louis, MO, USA). Phosphate-buffered saline solution was purchased from Biochrom (Berlin, Germany), and sodium chloride (NaCl) was acquired from VWR (Leuven, Belgium). Isoflurane (Isoflo®) was obtained from Abbott Animal Health (North Chicago, IL, USA). Harris haematoxylin was purchased from Harris Surgipath (Richmond, IL, USA), and 1% aqueous eosin from Australian Biostain (Traralgon, Australia). The Bio-Rad DC protein assay kit was obtained from Bio-Rad Laboratories (Hercules, CA, USA). Primary antibodies were acquired from different sources which are indicated in Supplementary Table S1 . Goat anti-rabbit IgG-horseradish peroxidase (ab97051) and goat anti-mouse IgG-horseradish peroxidase (ab6728) were provided by Abcam (Cambridge, UK), while enhanced chemiluminescence (Clarity Western ECL, 1.705.060) reagents and the Bio-Rad DC protein assay kit were purchased from Bio-Rad Laboratories (Hercules, CA, USA). Amersham Protran nitrocellulose blotting membranes (0.45 µm) were supplied by Cytiva (Buckinghamshire, UK).
7
Doxorubicin-based Cancer Treatment Protocol
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Doxorubicin hydrochloride (≥ 98% purity, DOX), Ponceau S, direct red 80, bovine serum albumin, and the all other chemicals used were obtained from Sigma-Aldrich (St. Louis, MO, USA). Phosphate buffered saline solution was purchased from Biochrom (Berlin, Germany), and sodium chloride (NaCl) was acquired from VWR (Leuven, Belgium). Isoflurane (Isoflo®) was acquired from Abbott Animal Health (North Chicago, IL, USA). Harris haematoxylin was purchased from Harris Surgipath (Richmond, IL, USA), and 1% aqueous eosin from Australian Biostain (Traralgon, Australia). The Bio-Rad DC protein assay kit was obtained from Bio-Rad Laboratories (Hercules, CA, USA). Primary and secondary antibodies used in Western blot and immunohistochemistry are shown in Supplementary Table S1. Enhanced chemiluminescence (Clarity Western ECL, ref. 1705060) reagents and the Bio-Rad DC protein assay kit were bought from Bio-Rad Laboratories (Hercules, CA, USA). Amersham Protran nitrocellulose blotting membranes (0.45 µm) were supplied by Cytiva (Buckinghamshire, UK).
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