Amiodarone (Sanofi K.K., Tokyo, Japan), veratrine (Santa Cruz Biotechnology, Dallas, TX), NS1643 (Abcam, Cambridge, England), and isoproterenol (Kowa Pharmaceutical, Tokyo, Japan) were dissolved or suspended in normal saline. BAYK8644 (Abcam, Cambridge, England) was dissolved in dimethyl sulfoxide and diluted with normal saline (the final concentration of dimethyl sulfoxide was 0.5%). All drugs were freshly prepared before use and administered intraperitoneally. The dose of Amiodarone (50 mg/kg) was decided based on the estimated body surface area of mice used in the present study (0.008 m2)13 (link) and the average body surface area of an adult human (1.86 m2),14 (link) and is considered equivalent to the clinical dosage for an adult human (300 mg). Other doses of Amiodarone (25, 100, and 200 mg/kg), and antagonists were chosen based on previously published data.15 (link)–19 (link)
Bay k 8644
Manufactured by Abcam
Sourced in United States
Bay K 8644 is a calcium channel agonist. It functions to activate L-type calcium channels, leading to an increase in calcium influx.
Automatically generated - may contain errors
Lab products found in correlation
Dmem f12, by Thermo Fisher Scientific (2 mentions)
Amiodarone, by Sanofi (1 mentions)
Lamotrigine, by Merck Group (1 mentions)
Ryanodine, by Merck Group (1 mentions)
Nifedipine, by Merck Group (1 mentions)
Cytosine β d arabinofuranoside ara c, by Merck Group (1 mentions)
Veratridine, by Abcam (1 mentions)
Cisapride, by Abcam (1 mentions)
Bovine serum albumin (bsa), by Thermo Fisher Scientific (1 mentions)
E 4031, by Abcam (1 mentions)
Triton x 100, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
4 aminopyridine, by Merck Group (1 mentions)
Alexa fluor 488 conjugated goat anti rabbit, by Thermo Fisher Scientific (1 mentions)
Quinidine, by Bio-Techne (1 mentions)
Verapamil, by Abcam (1 mentions)
Carbamazepine, by Merck Group (1 mentions)
Phenytoin, by Merck Group (1 mentions)
10 cm dish, by Corning (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
4 protocols using bay k 8644
1
Dosage and Formulation of Cardiovascular Drugs
2
Cardiac Ion Channel Pharmacology Assay
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Bay K 8644, veratridine, terfenadine, cisapride, anti-cTnT (1:400) antibody, E-4031 and verapamil were purchased from Abcam (Cambridge, MA, USA). Quinidine was from Tocris Bioscience (Bristol, UK). Nifedipine, 4-aminopyridine, fetal bovine serum (FBS), cytosine β-D-arabinofuranoside (ARA-C), poly-D-lysine and epinephrine, FLA-365, ryanodine, lamotrigine, carbamazepine, phenytoin and all inorganic salts were purchased from Sigma-Aldrich (St. Louis, MO, USA). Bovine serum albumin, DMEM/F12, Triton™ X-100, Alexa Fluor® 488-conjugated goat anti-rabbit were from ThermoFisher Scientific (Waltham, MA). The Ca2+ dye Fluo-8/AM was from TEFLab (Austin, TX, USA).
3
Isolation and Culture of BMMSCs
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Primary BMMSCs were isolated as previously described (Liao et al., 2016 ). Briefly, BMMSCs from 3‐month‐old mice were obtained and cultured in α‐MEM (Gibco BRL) supplemented with 20% fetal bovine serum (FBS; Thermo Electron), 0.292 mg/ml glutamine, 100 U/ml penicillin, and 100 mg/ml streptomycin (Invitrogen). Cells were then plated in the 10 cm dish (Costar) at 37°C in 5% CO2, and the medium was changed every 3 days. BMMSCs at passage 1 were used in the experiments. Bay K8644 (Abcam) was dissolved in DMSO (0.1% culture concentration).
4
Compound Screening in Cell Culture
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The compounds assayed in this study and their concentrations are listed in Table 1. Isoproterenol, propranolol, digoxin, astemizole, moxifloxacin, dl-sotalol, flecainide, terfenadine and verapamil were purchased from Sigma-Aldrich. E-4031 was purchased from Enzo Lifesciences (Farmingdale, NY, USA). Milrinone, forskolin, quinidine and aspirin were purchased from Wako Pure Chemical Industries (Osaka, Japan). Lidocaine was purchased from Nacalai Tesque (Kyoto, Japan). Bay K 8644 was purchased from Abcam (Cambridge, MA, USA). Chromanol 293B was purchased from Tocris Bioscience (Bristol, UK). All reagents were dissolved in dimethyl sulfoxide (DMSO) (Wako Pure Chemical Industries). The final concentration of each compound in the culture medium was at most 0.5% v/v. Each concentration was tested in triplicate or quadruplicate.
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