For β3-AR stimulation, mice were gavaged with 1 mg/kg CL 316,243 (C5976, Sigma) dissolved in water 3 hrs before analysis. For gavage of macronutrients 0.2 ml of 30% glucose (w/v) (G8270, Sigma), 30% BSA (w/v) (BAH62, Equitech-Bio) and 15% extra virgin olive oil (v/v) (Bertolli) was used as described before (Wang et al., 2016 (link)). Mice were gavaged once every hour for 4 hrs. For hexokinase inhibition mice were gavaged with 750 mg/kg 2-deoxyglucose (D6134, Sigma) or 170 mg/kg D-mannoheptulose (97318, Sigma) in water once every hour for 4 hrs. For pharmacological AMPK activation mice were gavaged with a single dose of 50 mg/kg phenformin HCl (S2542, Selleckchem) dissolved in water unless indicated otherwise, or 50 mg/kg A-769662 (S2697, Selleckhem) suspended in 1% carboxymethylcellulose (C4888, Sigma), and blood and bone marrow were analyzed 4 hrs later. Insulin (HumulinR, Lilly) was injected intraperitoneally into mice that fasted for 3 hrs at a dose of 1 unit/kg. For CCL2 reconstitution mice were fasted overnight and received 80 ug/kg CCL2 protein (479-JE/CF, R&D) in PBS intravenously 3 hrs before analysis. 4hr treatments were started at 9am (ZT2), 3hr treatments at 10am (ZT3) in the home cage and mice were analyzed at 1pm (ZT6).
C4888
Manufactured by Merck Group
Sourced in United States
C4888 is a laboratory instrument designed for specialized analytical applications. It is a high-precision, versatile tool that can be used to perform various types of analyses in a controlled and accurate manner. The core function of C4888 is to enable researchers and scientists to gather reliable data and insights within their respective fields of study.
Automatically generated - may contain errors
Lab products found in correlation
2 deoxyglucose d6134, by Merck Group (1 mentions)
G8270, by Merck Group (1 mentions)
479 je cf, by R&D Systems (1 mentions)
Humulin r, by Eli Lilly (1 mentions)
A6014, by Merck Group (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Glutamine, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
8 protocols using c4888
1
Pharmacological Modulation of Metabolic Pathways
2
Dye-Tagged Cellulose Film Fabrication
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First, 20 mg of acridine orange hemi (Zinc chloride) salt (A6014, Sigma Aldrich, St. Louis, MO, USA) dye was placed in 50 mL of distilled water and stirred for 60 min. In parallel, 2 g of carboxymethyl cellulose (CMC) sodium salt, medium viscosity (C4888, Sigma Aldrich, St. Louis, MO, USA) powder was thoroughly dispersed in 50 mL of distilled water and stirred for 60 min. After complete dispersion, the 50 mL of the CMC solution were added to the 50 mL of the dye solution. The whole mixture was stirred for 24 h at room temperature under dark conditions. Then, 10 mL of the CMC sodium salt tagged with dye solution was spread onto a petri dish. The solution was dried under a hood overnight, and a smooth film was obtained. All further investigations were performed on the films within the petri dishes.
3
Plaque Assay for Viral Titer Determination
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Viral titers of all the isolates were determined by plaque assay on BHK-21 cells. Briefly, 50,000 BHK-21 cells were seeded in each well of a 24 well plate. Virus was serially diluted 10-fold in assay diluent containing Minimum Essential medium (MEM) (Gibco-11090-073) containing 2% heat-inactivated fetal bovine serum (FBS) (Gibco-16140-071), 100 U/mL of penicillin, streptomycin and Glutamine (Gibco-10378-016) and BHK-21 cells were infected with serial dilutions of virus at 37 °C, 5% CO2 for 1 h on rocker. Post-infection inoculum was removed and cells were overlaid with 0.5% carboxymethylcellulose solution (CMC; Sigma – C4888) prepared in assay diluent as described above. The plates were incubated at 37 °C under a humidified atmosphere of 5% CO2 for 76 h for DENV-1 and 2, for 90 h for DENV-3, 80 h for DENV-4 and 48 h for ZIKV. After infection for the required time CMC solution was aspirated and cells were fixed with 3.7% formaldehyde solution for 30 min. Formaldehyde solution was washed with tap water and cells were stained with crystal violet solution for 10 min. Plates were washed with tap water and allowed to dry. Post-drying plaques were counted and titers represented as Plaque Forming Unit (PFU)/ml.
4
Cordyceps militaris Bioactive Compounds
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The commercially pulverized crude powders of the combined fruiting body and mycelium of Cordyceps militaris (CM), denoted as CmNo1 used in this study was provided by Liwanli Innovation Co., Ltd. (Taipei, Taiwan). CmNo1 contained 179.0 mg/g polysaccharides, 33.66 mg/g cordycepic acid, 0.91 mg/g adenosine, and 9.49 mg/g cordycepin. To determine the quantities of adenosine and codycepin present in CmNo1, the high-performance liquid chromatography (HPLC) DAD (Hitachi Co., Japan) analysis was performed while polysaccharide and cordycepic acid was quantified by UV-VIS spectroscopy. Thereafter, a suspension of CmNo1 was prepared with 2% carboxymethylcellulose (CMC; C4888, Sigma-Aldrich, USA) solution in water and administered orally to mice by stomach gavage at a dose of 360 mg/kg/day for 8 week.
5
Siponimod Ameliorates Experimental Autoimmune Encephalomyelitis
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Siponimod (BAF312; ADV638392161, Sigma-Aldrich) was suspended in 0.5% carboxymethylcellulose (CMC) medium viscosity (C4888, Sigma-Aldrich) and dosed adjusted to body weight (BW) with 3 mg/kg per day. This dosing was chosen because it has been shown to ameliorate EAE in rats, whereas a dose similar to that used in patients with MS (0.03 mg/kg BW) did not.21 (link) The vehicle alone, 0.5% CMC, was administered to a control group. In an attempt to prevent EAE (prevention paradigm), mice received the first dose at the age of 26 ± 2 days (n = 17 siponimod treated; n = 14 vehicle treated). For the evaluation of the therapeutic potential (treatment paradigm), mice were treated when reaching a clinical score of ≥3 (n = 7 siponimod treated; n = 8 vehicle treated). Considering the sex and the age when EAE started, animals were alternately assigned to treatment groups and received the respective agent daily via oral gavage. To minimize potential confounders, only mice of the same treatment group were held together in 1 cage. Mice were killed 30 days after the first application. For histologic reference at peak of disease, 5 mice were dissected when they reached a score of 3 without any further treatment.
6
Virus Titration in Hamster Lung Tissue
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Virus titration was performed on oropharyngeal swabs and lung tissue samples. The left lobe of the lung collected from euthanized hamsters were weighed and homogenized in 1 mL serum-free MEM using pestle. Tissue homogenates were clarified by centrifugation at 13,000 rpm for 10 min at 4 °C. Plaque assay was performed as described with some modifications [14] (link). Briefly, Vero cells (ATCC: CCL-81) grown in minimum essential medium (MEM, 61,100–061, (Thermo Fisher Gibco, Waltham, MA, USA) supplemented with 10% fetal bovine serum were seeded at a concentration of 3.5 × 105 cells/well on the day before the assay. Serial dilutions of each sample were added to the wells. The plate was incubated at 37 °C, 5% CO2 for 1 h and shaking the plates every 15 min. Then, the cells were overlaid with 1.5 mL/well of overlay medium containing MEM supplemented with 2% FBS and 1.5% carboxymethylcellulose (C4888, Sigma Aldrich, St. Louis, MO, USA). The culture was incubated at 37 °C, 5% CO2 for three days for plaque development. Then the overlay was removed, cell monolayers were fixed with 10% formaldehyde, and stained with 1% crystal violet. Viral titers were reported as PFU/g of sample.
7
Oral Delivery of PXL770 in CMC/Tween80
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PXL770 (synthesized as noted above) was suspended in 0.5% CMC/Tween80 (Sigma, #C4888, #P1754), and was administered twice a day by oral gavage at 75 mg/kg. Control animals received vehicle (0.5% Carboxy MethylCellulose - CMC) administered once a day.
8
Synthesis and Characterization of CMC
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CMC 2.24 was synthesized and provided by Chem-Master Intl. Inc. (99.5% pure, Stony Brook, NY, USA). Carboxymethyl cellulose as a vehicle was purchased from Sigma (C-4888, Sigma, USA). All cell culture reagents and chemical reagents were purchased from Thermo Fisher Scientific (Waltham, MA, USA).
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