Chapter 18 Purification of Muscle Actin

Publisher Summary This chapter focuses on the purification methods of muscle actin. With the advent of polyacrylamide gel electrophoresis as a highly resolving analytical tool for ascertaining protein purity, it became evident that muscle actin isolated by classic procedure contained significant amounts of actomyosin-associated muscle proteins such as tropomyosin and α-actinin. SDS-polyacrylamide gels have met with widespread use as a general method for obtaining muscle actin. A problem in establishing methods for actin purification resides in the level of purity. Emerging experimentation in cell biology and, specifically, cytoskeletal biochemistry requires probing sensitive properties of actin itself and actin associations with other cell proteins. The chapter explores some of the pitfalls associated with actin purification and clarifies in some detail the correct usage and expected result from each step of the widely used muscle actin purification. Additional steps to eliminate trace contaminants are also described.

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