The sediment was then resuspended at 1,000 × g for 15 min at 20 ± 1°C, and the supernatant was decanted. The sediment was again resuspended in 5 vol (vol/wt) of the original isolating medium and passed through a polyethylene strainer (0.1-mm pore size) to remove the connective tissue and debris. Five more volumes, resulting in 10 vol (vol/wt) total of the original isolating medium, were used to further facilitate passage of myofibrillar protein through the strainer. The supernatant was again sedimented at 1,000 × g for 15 min at 20 ± 1°C, after which it was decanted.