In the sum of 10 athymic female nude mice, 8-week-old mice with an average weight at 20 g were divided into two groups. 2,2,2-Tribromoethanol (2%, 200 μl/mouse; Sigma, St. Louis, MO, USA) was used to anesthetize the mice with intraperitoneal injection before implantation. A total of 1 × 106 K1 cells stably transfected with lncRNA FOXP4-AS1 were transferred subcutaneously in mouse flanks. Xenograft size was measured every other day. Then the final volume of xenografts was defined by V = 0.5 × L (length of tumor) × W2 (width of tumor). Forty-eight days later, all the treated nude mice were anesthetized to death and engrafted tumors were resected and their weight measured. Our study was approved and conducted by the Institutional Animal Care and Experimental Use Committee of Guizhou Medical University.
2 2 2 tribromoethanol
Manufactured by Merck Group
Sourced in United States, Germany, Japan, United Kingdom
2,2,2-tribromoethanol is a chemical compound used in research and laboratory settings. It is a colorless, crystalline solid with a boiling point of approximately 203°C. The compound is commonly used as a sedative and anesthetic agent in animal studies.
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Lab products found in correlation
2 methyl 2 butanol, by Merck Group (15 mentions)
Tert amyl alcohol, by Merck Group (9 mentions)
Avertin, by Merck Group (6 mentions)
Ethanol, by Merck Group (3 mentions)
Micron 4 retinal imaging microscope, by Phoenix Pharmaceuticals (3 mentions)
Sodium nitrite, by Merck Group (3 mentions)
Mayer s hematoxylin, by Merck Group (3 mentions)
Urethane, by Merck Group (3 mentions)
Methanol, by Merck Group (2 mentions)
Palm microbeam system, by Zeiss (2 mentions)
Tissue freezing medium, by Electron Microscopy Sciences (2 mentions)
1.0 pen membrane slides, by Zeiss (2 mentions)
Poly l lysine (pll), by Merck Group (2 mentions)
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Live dead baclight bacterial viability kit, by Thermo Fisher Scientific (2 mentions)
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133 protocols using 2 2 2 tribromoethanol
1
Xenograft Tumor Growth Dynamics
2
Acute Lung Injury Induction and PLAG Treatment
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Balb/c mice (9-week to 11-week-old males) were purchased from Koatech Co. (Pyongtaek, Republic of Korea) and maintained under specific pathogen-free (SPF) conditions. For the ALI model, mice were anesthetized with 2,2,2-Tribromoethanol (150 mg/kg; Sigma-Aldrich, St. Louis, MO, USA) by intraperitoneal injection and administered LPS intranasally (25 mg/kg; Sigma-Aldrich). PLAG (10, 50, or 250 mg/kg, Enzychem Lifesciences Co., Daejeon, Republic of Korea) was administered orally. Collection of bronchoalveolar lavage fluid (BALF) was performed by tracheal cannulation, using cold phosphate-buffered saline (PBS). Complete blood counts (CBCs) were performed using the Mindray BC-5300 Auto Hematology Analyzer (Shenzhen Mindray Bio-medical Electronics, China).
3
Syngeneic Islet Transplantation in Mice
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On the day of transplantation, 100 to 400 islets were manually counted and handpicked; islets with central necrosis and/or a damaged capsule were discarded. Animals were anesthetized with an intraperitoneal injection of Avertin (0.02 mL/g), a 2.5% solution of 10 g 97% 2.2.2-tribromoethanol (Sigma-Aldrich) in 10-mL 2-methyl-2-buthanol (Kemila, Stockholm, Sweden). In all animals (n = 12), 2 islet grafts were transplanted (n = 24 islet grafts, see Table 1 ). A midline incision was made in the skin, and the abdominal muscle was exposed. Syngeneic islets were infused into the muscle at both the left and right sides using a butterfly needle (25G). In each case, the infused volume was 100 to 120 μL.
4
Stereotaxic Delivery of Recombinant AAVs
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For stereotaxic delivery of recombinant AAVs, 9–week-old C57BL/6 N mice were anesthetized by inhalation of isoflurane (3–4%) or intraperitoneal injection of a saline solution containing 2% 2,2,2-tribromoethanol (Sigma), and secured in a stereotaxic apparatus. Viral solutions were injected with a Hamilton syringe using a Nanoliter 2010 Injector (World Precision Instruments) at a flow rate of 100 nl/min (injected volume, 0.6 μl). The coordinates used for stereotaxic injections into the hippocampal DG of mice were as follows: anteroposterior (AP), − 2.2 mm; medial–lateral (ML), ± 1.3 mm; and dorsal–ventral (DV), 2.2 mm from bregma. Each injected mouse was returned to its home cage and after 2 weeks was used for scoring seizure-like behaviors, immunohistochemical analyses, or electrophysiological recordings.
5
Echocardiographic and Electrocardiographic Analysis in Mice
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Echocardiographic and electrocardiographic analyses of mice were performed as described previously [27 (link),28 (link)]. After mice had been anesthetized with isoflurane (1.5–20% for maintenance; up to 5% for induction; Abbott Laboratories Pty Ltd.), echocardiography was performed in mice whose heart rate (HR) was between 500/min and 600/min using a Vevo 2100 system (VisualSonics) equipped with a 30-MHz microprobe. For electrocardiography, mice were anesthetized with 2,2,2-tribromoethanol (0.20 mg/g, Sigma-Aldrich) and electrocardiographic recordings were performed using CardiofaxVET ECG-1950 (Nihon Kohden, Tokyo, Japan).
6
Adipogenesis Regulation by Ginsenosides
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All reagents used in the experiment were guaranteed reagent grade and HPLC-grade. Acetonitrile, ethanol and methanol were from Merck (Darmstadt, Germany). Isopropanol (100%) was from J.T. Baker Chemical (Phillipsburg, NJ, USA). Phosphate-buffered saline (PBS) was from Lonza (Walkersville, MD, USA). Dulbecco’s modified Eagle’s medium (DMEM) was from BioWest (Riverside, MO, USA). Paraformaldehyde (4%) was from Biosesang (Seongnam-si, Korea). Bovine calf serum (BCS), fetal bovine serum (FBS), penicillin/streptomycin (P/S) and insulin were from Gibco (Grand Island, NY, USA). Ginsenoside Rg1, Rb1 and Rg3(S) were from ChromaDex Co. (Irvine, CA, USA). Dexamethasone (Dex), 3-isobutyl-1-methylxanthine (IBMX), thiazolyl blue tetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), Oil Red O, glycyrrhizin, 2-methyl-2-butanol and 2,2,2-tribromoethanol (Avertin) were from Sigma-Aldrich (St. Louis, MO, USA). RIPA buffer and bicinchoninic acid (BCA) protein assay kit were from Thermo Fisher Scientific (Waltham, MA, USA). Primary antibodies, anti-rabbit β-actin, PPARγ, C/EBPα, adiponectin, AMPK, p-AMPK, ACC, p-ACC were from Cell Signaling Technology (Danvers, MA, USA), SREBP-1c and CPT-1 from Santa Cruz Biotechnology (Dallas, TX, USA).
7
Isolation of Hippocampal Neurons from Mecp2 Mouse Model
8
IRBP-induced Murine Experimental Autoimmune Uveitis
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Female animals were used for all of the experiments otherwise mentioned. All animal experiments followed the guidelines of the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research and were approved by the Animal Care Committee of the Massachusetts Eye and Ear Infirmary. C57BL/6J mice (stock no. 00664) were purchased from Jackson Laboratories (Bar Harbor, ME). Standard laboratory chow was fed. Mice were allowed free access to food and water in a climate-controlled room with a 12-hour light/12-hour dark cycle. All mice used for experiments were 7-10 weeks old. For anesthesia, intraperitoneal injection of 250 mg/kg of 2,2,2-tribromoethanol (Sigma-Aldrich Corp., St. Louis, MO) was used for survival procedures, and 400 mg/kg was used for non-survival procedures. High performance liquid chromatography-purified human interphotoreceptor retinoid binding protein peptide 1- 20 (IRBP-p) was purchased from Biomatik (Wilmington, DE). Complete Freund’ s Adjuvant and Mycobacterium tuberculosis H37Ra were purchased from Difco (Detroit, MI).
9
Electrophysiological and Visual Acuity Assessments in Mice
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ERG measurements were done as described previously (Kim et al., 2017 (link)). In brief, mice were either dark- or light-adapted for 12 h and anesthetized with 2,2,2-tribromoethanol (Sigma, USA) in prior to dilating the pupils of the mice by 0.5% tropicamide. The mice were placed with a gold-plated objective lens on their corneas and silver-embedded needle electrodes at their foreheads and tails. The ERG recordings were performed using Micron IV retinal imaging microscope (Phoenix Research Labs, USA) and analyzed by Labscribe ERG software according to the manufacturer’s instruction.
Mouse visual acuity was measured with the OptoMotry system (Cerebral Mechanics, USA) as previously described (Prusky et al., 2004 (link)). Mice adapted to ambient light for 30 min were placed on the stimulus platform surrounded by four computer monitors displaying black and white vertical stripe patterns. An event that mice track the stripe movements with reflexive head-turn was counted as a successful visual detection. The detection thresholds were then obtained from the OptoMotry software.
Mouse visual acuity was measured with the OptoMotry system (Cerebral Mechanics, USA) as previously described (Prusky et al., 2004 (link)). Mice adapted to ambient light for 30 min were placed on the stimulus platform surrounded by four computer monitors displaying black and white vertical stripe patterns. An event that mice track the stripe movements with reflexive head-turn was counted as a successful visual detection. The detection thresholds were then obtained from the OptoMotry software.
10
UVB-Induced Skin Damage and PRE Treatment
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Five-week-old female albino hairless mice (SKH-1) were purchased from Orient Bio Inc. (Gyeonggi-do, Korea). The experimental protocols were approved by the Institutional Animal Care and Use Committee of Yonsei Laboratory Animal Research Center (Permit number: 201509-471-03). Eighteen mice were randomly divided into three groups: (1) normal group, (2) UVB group, and (3) UVB + PRE group. For 8 weeks, mice of the UVB and the UVB + PRE groups were exposed to UVB every alternate day using the UV crosslinker CL-1000M (UVP). The UVB dose started at 75 mJ/cm2. Then, the dose was increased by 1 minimal erythema dose (MED) per week, until 3 MED, which was maintained until end of the experiments. Mice of the UVB + PRE group were orally given 300 mg/kg/day PRE and the remaining groups received saline. After 8 weeks, the mice were anesthetized using 2,2,2-tribromoethanol (Sigma-Aldrich) and sacrificed. For optical microscopy, the dorsal skin samples were fixed in 10% formalin and the residual dorsal skin was stored at −80°C.
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