A method for the preparation of skeletal muscle sarcolemma

This procedure releases the trapped sarcolemmae from the gel and simultaneously liberates the remaining intracellular contents, by treating the gel with ATP and then quickly washing in aqueous solutions.

Abstract

A new procedure is described for obtaining the sarcolemma of striated muscle. In essence the technique utilizes: (a) shredding and shearing of striated muscle fibers in a reverse-bladed Waring blendor in a solution of 50 mM CaCl2 at 0°, (b) filtration through plastic screening at 0° to remove the fibrous connective tissue, (c) incubation at 37° in dilute salt solutions designed to disrupt internal structure, (d) washing procedures designed to evacuate the intracellular components and separate them from the sarcolemma (Figs. 1, 2), and (e) dissolution of the actomyosin gel, resulting from the breakdown of the myofibrils (Fig. 4), by treating the gel with ATP and then quickly washing in aqueous solutions. This procedure releases the trapped sarcolemmae from the gel and simultaneously liberates the remaining intracellular contents.