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13 protocols using histamine

1

Cellular Signaling Pathway Assay

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DMEM, FBS, penicillin/streptomycin, L-glutamine, and trypsin-EDTA were purchased from Capricorn Scientific, NEBuilder HiFi DNA Assembly kit and Q5 Polymerase from New England Biolabs, Effectene Transfection Reagent from Qiagen, a saponin from AppliChem, poly-L-lysine hydrobromide, GTPγS, acetylcholine, and carbachol from Sigma-Aldrich, histamine from Alfa Aesar and pertussis toxin from Merck.
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2

Synthesis of Molecularly Imprinted Polymers

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The monomers used for the synthesis of the MIP- and NIP-particles, ethylene glycol dimethacrylate (EGDM), methacrylic acid (MAA), acrylic acid (AA) and acrylamide (AM), were purchased from Acros (Geel, Belgium). For EGDM, MAA and AA the stabilizers were removed by filtration over alumina powder prior to polymerization. The solvent dimethylsulfoxide (DMSO), serving as a porogen was obtained from Acros and the azobisisobutyronitrile (AIBN) initiator was acquired from Fluka (Buchs, Switzerland). The template molecules histamine and serotonin were obtained from Alfa Aesar (Karlsruhe, Germany). l-Nicotine was purchased from Acros. All solvents were of analytical grade.
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3

In Vitro Electrochemical Sensing Setup

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Adenosine, dopamine, histamine, and Adenosine triphosphate (ATP) were purchased from Acros Organics (Morris Plains, NJ), and H2O2 was purchased from Macron (Center Valley, PA). Stock solutions were prepared in 0.1 M HClO4 to 10 mM concentration. The final working solutions were prepared by diluting the stock solution in the phosphate-buffered saline (PBS) containing 131.25 mM NaCl, 3.00 mM KCl, 10.0 mM NaH2PO4, 1.2 mM MgCl2, 2.0 mM Na2SO4, and 1.2 mM CaCl2 with pH adjusted to 7.4. The buffer was also adjusted to pH 7.3 by HCl or pH 7.5 by NaOH to test the effect of pH change. In vitro experiments and electrode calibration were conducted with a flow cell connected to a syringe pump (Harvard Apparatus, Holliston, MA) and a six-port loop injector with an air actuator (VIVI Valco Instruments, Houston, TX).
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4

Electrochemical Sensor for Histamine Detection

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Potassium nitrate (KNO3), hydrogen peroxide 35% (w/v), and histamine 97% (w/w) were purchased from Acros organics (Springfield Township, NJ, USA). Potassium ferrocyanide trihydrate (K3Fe(CN)6) was acquired from HiMedia Laboratories (Mumbai, India). Phosphate buffer saline (PBS 1X) was prepared with sodium phosphate dibasic (NaHPO4), potassium phosphate (KH2PO4), potassium chloride (KCl), and sodium chloride (NaCl), obtained from Chem Center (La Jolla, CA, USA). Copper sulfate (23.5 wt %, as fertilizer) was acquired from an agricultural supply store (Campofert S.A.S., Yumbo, Colombia).
Purified diamine oxidase (DAO) from porcine kidney was obtained from Sigma Aldrich (St. Louis, MO, USA). DAO (7 wt % as dietary supplement) was obtained from Swanson Health Products, Inc. (Fargo, ND, USA). The formulation contained microcrystalline cellulose, sucrose, ascorbic acid, rice starch, hydrated magnesium silicate, corn starch, carboxymethylcellulose, titanium dioxide, and glycerol (proprietary concentrations). Kapton (polyimide) tape, of approximately 30.4 μm film thickness and 50 mm width, was obtained from McMaster-Carr Co. (Elmhurst, IL, USA), silver/silver chloride ink for screen printing was acquired from DuPont (Wilmington, DE, USA). Nine volt batteries were acquired from a local hardware store.
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5

HUVEC Isolation and Activation Protocol

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Human umbilical vein endothelial cells (HUVEC) were isolated from acquired umbilical cords using a collagenase perfusion technique. HUVEC were grown in T75 flasks at 37 °C and 5% CO2 until confluent and then seeded at a confluent density onto glutaraldehyde-crosslinked gelatin coated 30 mm glass round coverslips (Warner Instruments). Coverslips were utilized 36–48 h after seeding.
HUVEC were activated with 1 ng/mL IL-1β (Fitzgerald) in cell media 4 h before use in flow experiments. HUVEC were also activated with 100 µM histamine (Acros Organics) 2 min before use in flow experiments.
For E-selectin blocking experiments, activated HUVEC were blocked with 20 µg/mL anti-E-selectin (R&D Systems #BBA26) or IgG1 isotype control (BioLegend #400102) for 30 min before use in flow experiments to prevent leukocyte adhesion.
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6

Histidine Metabolism Pathway Analysis

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Water (18.2 MΩ cm À1 ) was purified with a Milli-Q Plus ® system (Millipore, Madrid, Spain). The LC-MS solvent (acetonitrile, ACN) and additive (formic acid) were of UHPLC-MS-grade and were from Scharlau (Scharlab S.L., Barcelona, Spain). L-glutamate, imidazole-4-acetate, N-acetylhistamine, cis-urocanate, imidazole propionate and 2-amino-4-thiazoleacetic acid were from Sigma-Aldrich Chemie GmbH. Trans-urocanate, histamine, and atenolol were obtained from Acros Organics (Geel, Belgium).
F I G U R E 1 Metabolic pathways involving measured metabolites from the histidine metabolism L-histidine was from VWR International Ltd (UK). Other chemicals used for measuring enzymatic activities were BugBuster Protein Extraction Reagent (Novagen, Darmstadt, Germany), magnesium chloride, reduced glutathione, pyridoxal-5 0 -phosphate and aminoguanidine (Sigma-Aldrich, Chemie GmbH).
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7

Intracellular Calcium Assay for AsPC-1 Cells

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Intracellular
calcium concentration
of AsPC-1 cells was measured using the FLIPR-4 calcium assay reagent
(Cat # R8142, Molecular Devices, San Jose, CA). In brief, cells were
plated in black-walled clear-bottom 96-well plates overnight at ∼80%
confluency using media and conditions described in Methods. Culture media was then removed and cells were incubated
for 1 h at 37 °C, 5% CO2 in FLIPR-4 loading buffer,
consisting of FLIPR-4 reagent diluted (as per the manufacturer’s
instructions) in Hank’s balanced salt solution (HBSS) (with
calcium and magnesium) buffered with 20 mM N-(2-hydroxyethyl)piperazine-N′-ethanesulfonic acid and 0.2% bovine serum albumin,
with pH adjusted to 7.4. Loading buffer also contained probenecid
(2.50 mM, Sigma-Aldrich, Cat # P8761) to prevent leakage of the calcium
reagent from the cells. Calcium response was then measured via a FlexStation
3 Multi-Mode Microplate Reader (Molecular Devices). GPCR agonists
were added, and response in relative fluorescence units was measured
over 105 s for each well, yielding data for peak response and kinetics
of response. For calcium assays and wound-healing assays, we used
the following GPCR agonists: Neurotensin (Cat # 1909, Tocris, Minneapolis,
MN); 2-Thio-UTP (Cat # 3280, Tocris); histamine (Cat # AAJ6172703,
Fischer Scientific); oxytocin (Cat # 1910, Tocris); and sulprostone
(Cat # 14765, Cayman Chemical).
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8

Intracellular Calcium Signaling Assay

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Cell culture chemicals were obtained from Thermo Fisher Scientific; histamine, amphotericin B (AmB), KCl, MgCl2, CaCl2, glucose, HEPES, EGTA, 2-aminoethoxydiphenyl borate (2-APB), and ouabain from Sigma-Aldrich, NaCl from Carl Roth. KB-R7943 was purchased from Sigma-Aldrich and Abcam. We used Fura-2 AM products by both Sigma-Aldrich and Thermo Fisher Scientific.
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9

Measuring Gastric pH Under Histamine Stimulation

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Gastric pH was measured as described39 (link), 40 (link). In brief, gastroesophageal and gastroduodenal junctions were ligated and total gastrectomy was performed. Saline (500 μl) was injected into the lumen and the whole stomach was placed in an oxygenated bath (37 °C) containing HEPES buffer (pH 7.4). Histamine (Thermo Fisher Scientific; 200 μM) was added to the bath for 60 min, the injected non-buffered saline solution was aspirated from the stomach, and the pH was measured with a pH probe (Accumet AB15 Basic and BioBasic; Thermo Fisher Scientific).
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10

Comprehensive Reagent Acquisition Protocol

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Pyrogallol solution and Alcian blue were purchased from Oxford Labs (India). Elman’s reagent and Alizarin red S were purchased from Omicron Sciences Limited (United Kingdom). Follin’s reagent, carboxymethylcellulose, picric acid, EDTA, N-1-naphthyl ethylene amine dihydrochloride, sulfanilamide, thiobarbituric acid, sodium-potassium tartrate, DTNB, and Griess reagent were purchased from Sigma-Aldrich, United States. Histamine and testosterone were purchased from ThermoFisher Scientific, United States. All chemicals and solvents used in the study were of analytical or pharmaceutical grade.
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