Caco2 cells were obtained from the American Type Culture Collection (ATCC, Manassas VA) and YAMC cells were kindly donated by Dr. Robert Whitehead (Vanderbilt University). The corresponding knockout Caco2-AhRKO and YAMC-AhRKO cell lines were generated by CRISPR/Cas9 as described (29 (link),30 (link)). Caco2 cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 20% fetal bovine serum (FBS), 1 x MEM non-essential amino acid solution (Gibco), and 1 x antibiotic/antimycotic reagent (Sigma-Aldrich, St. Louis, MO) at 37°C in the presence of 5% CO2. YAMC cells were cultured in RPMI1640 media supplemented with 5% FBS, 1% glutamine, 0.1% ITS (Corning) and 10U IFNγ (Sigma-Aldrich) at 33°C in 5% CO2. The β-carboline alkaloids norharman, harmine hydrochloride, harmane, harmaline hydrochloride dehydrate, 1-methyl-2, 3, 4, 9-tetrahydro-1H-β-carboline-1-carboxylic acid sodium butyrate, sodium propionate and sodium acetate were purchased from Sigma-Aldrich. TCDD (99%) was synthesized in the Safe laboratory and roasted coffee (Seattle’s Best #4) was purchased locally.
Harmaline hydrochloride dehydrate
Manufactured by Merck Group
Sourced in United States
Harmaline hydrochloride dehydrate is a chemical compound used as a laboratory reagent. It is a crystalline solid with the chemical formula C₁₃H₁₄N₂O • HCl • 2H₂O.
Automatically generated - may contain errors
Lab products found in correlation
Harmane, by Merck Group (1 mentions)
Sodium propionate, by Merck Group (1 mentions)
Caco 2 cells, by Thermo Fisher Scientific (1 mentions)
Antibiotic antimycotic reagent, by Merck Group (1 mentions)
Norharman, by Merck Group (1 mentions)
Sodium acetate, by Merck Group (1 mentions)
Mem non essential amino acid solution, by Thermo Fisher Scientific (1 mentions)
Ifn γ, by Merck Group (1 mentions)
Caco 2 cell, by American Type Culture Collection (1 mentions)
Harmine hydrochloride, by Merck Group (1 mentions)
Matlab 2014a software, by MathWorks (1 mentions)
Adxl353, by Analog Devices (1 mentions)
2 protocols using harmaline hydrochloride dehydrate
1
Caco2 and YAMC Cell Line Generation
2
Harmaline-Induced Tremor Assessment in Rats
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On experiment days, rats received an intraperitoneal injection of harmaline solution (15 mg/kg) prepared by dissolving harmaline hydrochloride dehydrate (≥ 95%), (Sigma-Aldrich, St. Louis, MO, USA) in saline. Approximately 45 minutes after harmaline injection, the rats exhibited a whole body visible tremor that was recorded using a tri-axial accelerometer (ADXL353, Analog Devices, MA, USA). The accelerometer was mounted on the rats' back and secured with a hook-and-loop tape, which was sufficient to detect and measure the harmaline induced general body tremor. It was also a more comfortable and secure location as it does not affect the rats' movement and they cannot reach to bite it. The accelerometer data (three axes) were digitized using a digital analog converter (DAC, NI USB-6343, National Instruments, TX, USA) at 30 kHz sample rate, displayed and recorded for off-line analysis using a custom written MATLAB 2014a software (MathWorks, MA, USA).
When rats were laying still there was never tremor visible. Thus, for the experiment they were placed in a freely rotating wheel to ensure they were always moving, and the tremor was as consistent as possible.
When rats were laying still there was never tremor visible. Thus, for the experiment they were placed in a freely rotating wheel to ensure they were always moving, and the tremor was as consistent as possible.
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