Extraction of Total Protein from Cardiomyocytes and Western Blotting Analysis

Using total protein lysates for Western blotting confers greater benefits than traditional approaches that use fractionated samples, where a portion of the sample is discarded during processing. By incorporating low-volume inputs, it is possible to identify any protein present in the sample where a suitable antibody is available. The inclusion of a calibration curve allows quantitative analyses to be undertaken. There are additional benefits including that only requiring a small input saves on precious samples. Using whole muscle lysates is necessary in order to ascertain specificity of antibodies for subsequent immunofluorescent methods.