A concentrated ethanol extract of aronia berry was provided by POLA Chemical Industries Inc. (Kanagawa, Japan). According to a high-performance liquid chromatography (HPLC) analysis conducted by the manufacturer, the total anthocyanins content of the aronia berry extract powder was 36.49%, which consisted of 23.52% cyanidin-3-O-galactoside, 10.48% cyanidin-3-O-arabinoside, 1.36% cyanidin-3-O-xyloside, and 1.13% cyanidin-3-O-glucoside. The powder extract was dissolved in deionized water at 5 mg/mL and used in the experiments. RPMI-1640 medium, Hank’s balanced salts solution (HBSS), and 2’,7’-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Fetal bovine serum (FBS), penicillin/streptomycin, novoheparin, and recombinant human fibroblast growth factor (hFGF) basic were obtained from Biowest (Nuaillé, France), GIBCO (Grand Island, NY), Mochida Pharmaceutical (Tokyo), and R&D Systems (Minneapolis, MN), respectively.
Hank s balanced salts solution hbss
Manufactured by Merck Group
Sourced in United States
Hank's Balanced Salt Solution (HBSS) is a sterile, balanced salt solution commonly used in cell culture and other biological applications. It is formulated to maintain the pH, osmotic pressure, and ion concentrations required for the survival and growth of cells in vitro. HBSS does not contain any additional nutrients, growth factors, or serum components.
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Lab products found in correlation
Hanks balanced salt solution (hbss), by Merck Group (3 mentions)
Rpmi 1640 medium, by Merck Group (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Biowest (1 mentions)
Penicillin streptomycin, by Biowest (1 mentions)
Rpmi 1640 culture medium, by Thermo Fisher Scientific (1 mentions)
Histopaque 1077, by Merck Group (1 mentions)
Collagenase p, by Roche (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Phorbol 12 myristate 13 acetate pma, by Merck Group (1 mentions)
2 7 dichlorofluorescein diacetate dcfh da, by Merck Group (1 mentions)
Nmri mice, by Taconic Biosciences (1 mentions)
β mercaptoethanol, by Merck Group (1 mentions)
Serotonin creatinine sulfate, by Merck Group (1 mentions)
Imipramine hydrochloride, by Merck Group (1 mentions)
Citalopram hydrobromide, by Merck Group (1 mentions)
Pargyline hydrochloride, by Cayman Chemical (1 mentions)
3 4 methylenedioxymethamphetamine hydrochloride, by Merck Group (1 mentions)
8 protocols using hank s balanced salts solution hbss
1
Aronia Berry Extract Characterization
2
Neonatal Mouse Islet Isolation
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Protocols for islet isolation from neonatal mouse pancreata were modified from previously reported protocols developed in our laboratory [22 (link),23 (link)]. In brief, the dissected pancreata from five neonatal mice from each treatment group were pooled together for islet isolation. They were minced finely and digested in 3 mg/ml collagenase P (Roche Diagnostic, Indianapolis, IN) in 5 ml of Hanks balanced salts solution (HBSS) (Sigma-Aldrich, St. Louis, MO) for 7–10 min at 37°C with vigorous shaking. The digestion was terminated by addition of ice-cold HBSS followed by centrifugation for 5 min at 1600 rpm twice, and pancreatic islet tissues were then separated by Histopaque-1077 (Sigma-Aldrich) gradient density centrifugation. The re-suspended islets were handpicked under a stereomicroscope and then cultured in RPMI1640 culture medium supplemented with 10% fetal bovine serum, 1% penicillin, and streptomycin (Invitrogen, Carlsbad, CA) for 3 days before being subjected to functional assessment.
3
Terminalia bellirica Extract Bioactivity
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A hot water extract of Terminalia bellirica was provided by Toyo Shinyaku Co., Ltd. (Saga, Japan). The powder was dissolved in deionized water and used in the experiments. The contents of gallic acid and ellagic acid in the TBE stock solution (40 mg/mL) were 4.6 mg/mL and 0.16 mg/mL, respectively. RPMI-1640 medium, phorbol-12-myristate-13-acetate (PMA), Hank’s balanced salts solution (HBSS), and 2′,7′-dichlorofluorescein-diacetate (DCFH-DA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Fetal bovine serum (FBS) and penicillin/streptomycin were obtained from GIBCO (Life Technologies, Grand Island, NY, USA).
4
Isolation of Pancreatic Islets from Mice
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Pancreatic islets were isolated from adult female NMRI mice (Taconic) weighing 22‐25 g by collagenase digestion technique, as described previously.21 , 22 , 23 In brief, the mice were anaesthetized with pentobarbital intraperitoneally (50 mg/kg). A midline laparotomy was applied, and the distal end of the common bile duct was clamped at the papilla Vateri, whereafter the hepatic duct was cannulated and ~3 mL of ice‐cold Hanks’ Balanced Salts Solution (HBSS; Sigma Chemical) containing 0.3 mg/mL of Collagenase P (Boehringer Mannheim GmbH) was injected into the duct system of the pancreas. The pancreas was subsequently removed and placed in a test tube in water bath at 37°C for 19 minutes. After being rinsed for 3 times with HBSS, the islets were hand‐picked under a stereomicroscope and immediately transferred for an overnight incubation in PRMI 1640 medium. The medium contained 11 mmol/L glucose and supplemented with 10% foetal calf serum, 5 μmol/L β‐mercaptoethanol (Sigma‐Aldrich), 100 IU/mL penicillin and 100 μg/mL streptomycin (all obtained from GIBCO BRL, Paisley, UK, if not stated otherwise). The incubation condition was 37°C and 95% normal atmosphere/5% CO2. Islets for static incubation and perifusion studies were obtained from 12 to 20 mice to compensate for interindividual differences.
5
Radioactive Serotonin Binding Assay
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Tritium-labeled Serotonin creatinine sulfate (3H-serotonin; 28.3 Ci mmol−1 and 41.3 Ci mmol−1) and radiocarbon-labeled serotonin binoxalate (14C-serotonin; 54.0 mCi mmol−1) were obtained from Perkin Elmer (Waltham, MA, USA). Serotonin creatinine sulfate, imipramine hydrochloride, citalopram hydrobromide, 3,4-methylenedioxymethamphetamine hydrochloride (MDMA, “Ecstasy”), L-ascorbic acid, and Hanks’ Balanced Salts Solution (HBSS) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Pargyline hydrochloride was from Cayman Chemical (Ann Arbor, MI, USA). Other drugs were generous gifts from manufacturers.
6
Evaluating Drug Transport Mechanisms
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Phenformin, verapamil, cyclosporine A (CsA), propranolol, and Hank’s Balanced Salts Solution (HBSS) were purchased from Sigma-Aldrich (St. Louis, MO, USA). [3H]digoxin (1.103 TBq/mmol) was purchased from Perkin Elmer Inc. (Boston, MA, USA). Fetal bovine serum (FBS), Medium 199, gentamycin, hygromycin, and trypsin-EDTA were purchased from Hy-clone Laboratories. All other chemicals were of reagent grade.
7
Hippocampal Neuron Culture Protocol
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Hippocampal cultures were obtained as described (Piedras-Rentería et al., 2004 (link)). In brief, whole hippocampi were dissected from newborn rats in Hanks’ Balanced Salts Solution (HBSS) (Sigma, St. Louis, MO) supplemented with 20% of Fetal Bovine Serum (Cellgro, Herndon, VA) (20% FBS-HBSS). Hippocampi were washed with 20% FBS-HBSS and HBSS and incubated for 10 min at 37°C with trypsin type XI (3.4 mg/ml) (Sigma, St. Louis, MO) plus DNAse type I (600 U/ml) (Calbiochem, Billerica, MA). After digestion, the tissue was washed with 20% FBS-HBSS and HBSS and mechanically dissociated in HBSS supplemented with 600 U/ml DNAse I and 12 mM MgSO4. Cells were plated onto matrigel-coated coverslips (12 mm, Carolina Biological Supply, NC) at a density of 25,000–35,000 and kept in a 5% CO2 humidified atmosphere at 37°C. The media was supplemented with thymidine -β-D-arabinofuranoside (4 μM) after the second day in culture (2 DIV, days in vitro).
8
Cell Membrane Permeability Assay in T. cruzi
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The action of compounds 1d , 1e , 3d and 4a in the cell membrane permeability were evaluated in T. cruzi trypomastigotes (2 × 106/well) seeded in 96-well black polystyrene microplates. Parasites were washed and incubated in the dark with 1 μM SYTOX Green probe (Molecular Probes) in HANKS’ balanced salts solution (HBSS; Sigma-Aldrich) supplemented with 10 mM D-Glucose (HBSS + Glu) in 96-well black polystyrene microplates43 (link),44 (link). Each compound was added (t = 0 min) at IC50 concentration and fluorescence levels were measured every 20 min for up to 120 min. The maximum permeabilization was obtained with the addiction of 0.5% Triton X-100. Fluorescence intensities were determined using a fluorimetric microplate reader (FilterMax F5 Multi-Mode Microplate Reader-Molecular Devices) with excitation and emission wavelengths of 485 and 520 nm, respectively. The following internal controls were used in the evaluation: i) the background fluorescence of the compound at the respective wavelengths, ii) the possible interference of DMSO.
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