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Catalog Antibody | A primary hERG antibody (Sigma-Aldrich) and anti-rabbit HRP-conjugated secondary antibody (GenScript, Piscataway, New Jersey, USA) were used accordingly. To determine equal protein loading, β-actin (Santa Cruz Biotechnology Inc., Dallas, Texas, USA) was used. To improve the color, sharpness, and contrast of western blot images, the Format Picture setting in Microsoft Word 2010 was used. Graphical representation of hERG protein expression was quantified using Image J (http://imagej.nih.gov/ij/NIH) and normalized to the β-actin loading control | Get A Quote |
Background: Methadone is now increasingly being used to control opioid withdrawal symptoms, and recently, chronic pain. With increasing reports of cardiac problems induced by methadone, we aimed to determine effect of methadone on hERG expression after chronic treatment with racemic and the two enantiomers (S- and R-) of methadone. The molecular mechanism underlying methadone-disrupted hERG expression in human primary cardiomyocytes was also investigated. Methods: Cardiomyocytes were treated for 5 days with racemic-, (S)- and (R)-methadone at 3, 2, and 7 µM, respectively. The effect of racemic-, (S)-, and (R)-methadone on hERG, SGK3, and Hsp90 mRNA expression was examined by real-time PCR. Protein expression o... More