α-Actinin protein (AT01)

Material
α-actinin has been purified from rabbit skeletal muscle. The protein is supplied as a lyophilized powder in medium salt buffer (50 mM NaCl, 20 mM PIPES pH 7.2). The lyophilized product is stable at -70°C for at least 2 years. AT01 should be resuspended in nanopure water containing 1 mM β-mercaptoethanol or DTT to the desired working concentration.

Purity
Purity is determined by scanning densitometry of the protein on an SDS-PAGE gel. α-actinin is >90% pure.

Biological Activity
At 2.5 µM (0.25 mg/ml) α-actinin and 25 µM (1.0 mg/ml) F-actin at pH 7.0 and 24°C, approximately 70% of the α-actinin will co-sediment with F-actin after centrifugation at 150,000 x g for 1 h.

Examples of publications where this product was used:
Karakesisoglou, I., Yang, Y. and Fuchs, E. (2000). An epidermal plakin that integrates actin and microtubule networks at cellular junctions. J. Cell Biol. 149, 195-208.

Lu, S., Carroll, S. L., Herrera, A. H., Ozanne, B. and Horowits, R. (2003). New N-RAP-binding partners α-actinin, filamin and Krp1 detected by yeast two-hybrid screening: implications for myofibril assembly. J. Cell Sci. 116, 2169-2178.

Maul, R. S., Song, Y., Amann, K. J., Gerbin, S. C., Pollard, T. D. and Chang, D. D. (2003). EPLIN regulates actin dynamics by cross-linking and stabilizing filaments. J. Cell Biol. 160, 399-407.

van der Gucht, J., Paluch, E., Plastino, J. and Sykes, C. (2005). Stress release drives symmetry breaking for actin-based movement. Proc. Natl. Acad. Sci. U. S. A. 102, 7847-7852.

Waterman-Storer, C., Duey, D. Y., Weber, K. L., Keech, J., Cheney, R. E., Salmon, E. D. and Bement, W. M. (2000). Microtubules remodel actomyosin networks in Xenopus egg extracts via two mechanisms of F-actin transport. J. Cell Biol. 150, 361-376.