PMC:3824683 / 33786-34886 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/3824683","sourcedb":"PMC","sourceid":"3824683","source_url":"http://www.ncbi.nlm.nih.gov/pmc/3824683","text":"Western blotting\nThe protein levels were measured by analyses of western blots using whole-cell lysates obtained from astrocytes treated with appropriate reagents. Briefly, the cells were lysed using radioimmunoprecipitation assay (RIPA) buffer (Boston BioProducts, Ashland, MA, USA) supplemented with HaltTM Protease Inhibitor Cocktail (Pierce Biotechnology, Rockford, IL, USA). The protein concentrations in the lysates were measured using BCA Protein Assay Kit (Pierce Biotechnology). The protein samples (30 μg) were electrophoresed on a 10% polyacrylamide gel and transferred to PVDF membrane. Finally, the membrane was probed with appropriate primary and secondary antibodies for caspase-3, ferritin heavy chain (Ferritin HC), NOX2, NOX4, CYP2B6, CYP2E1, and CYP2D6 to measure their expression levels. The bands were detected using BM Chemiluminescence Western Blotting Substrate (POD) (Roche Applied Sciences, Indianapolis, IN, USA). The bands were analyzed using FlourChem HD2 software (Alpha Innotech, San Leandro, CA, USA) and the intensities were normalized using GAPDH as loading control.","divisions":[{"label":"Title","span":{"begin":0,"end":16}}],"tracks":[]}