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.

Fiber Type Identification of Human Skeletal Muscle.

The technique described here can be used to identify specific myosin heavy chain (MHC) isoforms in segments of individual muscle fibers using dot blotting, hereafter referred to as Myosin heavy chain detection by Dot Blotting for IDentification of muscle fiber type (MyDoBID). This protocol describes the process of freeze-drying human skeletal muscle and isolating segments of single muscle fibers. Using MyDoBID, type I and II fibers are classified with MHCI- and IIa-specific antibodies, respectively.