Mechanical Hysteresis in Actin Networks: Supplementary information
نویسندگان
چکیده
Monomeric actin (G-actin) was purified using protocol adapted from Ref.[1] from rabbit skeletal muscle acetone powder (Pel Freeze Biologicals, Product code: 41008-3). Small aliquots of actin solution were drop frozen in liquid nitrogen, and stored at -80 oC. For each experiment fresh aliquots of frozen actin were thawed and used. FLN was purified from chicken gizzard and aliquots were also stored at -80 oC. In-vitro network formation 10X actin polymerization-buffer (2 mM TrisHCl, 2 mM MgCl2, 100 mM KCl, 0.2 mM DTT, 0.2 mM CaCl2, 0.5 mM ATP, pH 7.5) was mixed with freshly thawed FLN. Freshly thawed G-actin was mixed with 1X Ca-G-buffer (2mM Tris-HCl, 0.2mM ATP, 0.5mM DDT, 1mM NaN2 and 0.1mM CaCl2, pH 8). 1X Ca-G-buffer containing G-actin and 10X actin polymerization buffer containing FLN were mixed gently for 10 s just before loading the sample to initiate the polymerization process of actin filaments in the rheometer sample chamber.
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