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2 protocols using nebivolol

1

Receptor Binding Assay Protocol

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Carvedilol was purchased from TCI America (Portland, OR). 4-(3-t-butylamino-2-hydroxypropoxy)-[5,7-3H]benzimidazole-2-one (3H-CGP) with a specific activity of 41.7 Ci/mmol was purchased from Perkin Elmer (Waltham, MA). Isoproterenol, 3-isobutyl-1-methylxanthine (IBMX), and forskolin were purchased from Sigma (St. Louis, MO). Atenolol, nebivolol, ICI-118,551, L-748,337, xamoterol humifumerate, formoterol humifumerate and L-755,507 were obtained from Tocris (Bristol, United Kindom). EGF was purchased from Peprotech (Rocky Hill, NJ) and dissolved in sterile deionized water as 100 ug/mL stock and stored in −20°C freezer.
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2

Screening of Small Molecule Modulators

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Acebutolol, alprenolol, atenolol, bucindolol, carazolol, carvedilol, CGP12177, ICI 118551, isoproterenol, labetalol, metoprolol, nadolol, nebivolol, pronethalol, timolol, and 2-{[β-(4-Hydroxyphenyl)ethyl]aminomethyl}-1-tetralone hydrochloride (HEAT HCl) were purchased from Tocris (Bristol, United Kindom). Propranolol HCl and 4-hydroxycarbazole were obtained from Sigma-Aldrich (St. Louis, MO) and bupranolol was obtained from Abcam (Cambridge, UK). All compounds were dissolved in DMSO to obtain a 10 mM stock concentration, which was stored at -20°C. EGF was purchased from Peprotech (Rocky Hill, NJ) and dissolved in sterile deionized water at a 10 ug/mL stock and stored at -80°C. Primers were purchased from IDT (Coralville, IA). The lentiviral vector pLV-H1-EF1α-puro, annealing buffer, packaging vector, and polybrene were purchased from Biosettia Inc. (San Diego, CA). 4-hydroxycarbazole (4-OHC) was purchased from Sigma Aldrich
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