Acetone Powders

…To isolate actin, 3 g of rabbit muscle acetone powder (Pel Freez Biologicals) was mixed with 100 mL of actin extraction buffer (2 mM Tris-HCl, 0.2 mM CaCl 2 , 0.2 mM ATP, and 1 mM…

…Nature Structural & Molecular BioloGy | www.nature.com/nsmb Articles Nature Structural & Molecular Biology Methods Protein purification. Rabbit skeletal muscle actin was extracted from acetone powder (Pel Freeze, catalog no. 41995) by stirring in G-buffer (20 ml g–1, 2 mM Tris pH 8.0, 0.2 mM ATP, 0.5 mM DTT, 0.1 mM CaCl2) on ice for 30 min before spinning in a Beckman JA-20 rotor at 16,000 r.p.m. for 30 min at 4…

…esults demonstrate a versatile approach to construct tunable, anisotropic macromolecular liquids. … Monomeric actin (G-actin) is purified from rabbit skeletal muscle acetone powder (Pel Freeze Biologicals, Product code: 41008-3) using a procedure adapted from ref. 27 and stored in 2…

…Purified protein was stored in HEPES buffer at 4 ◦C (20 mM HEPES, pH 7.5, 5 mM dithiothreitol (DTT) and 50 mM NaCl) and used within 2–4 weeks. Actin was purified from rabbit muscle acetone powder (Pel- Freez Biologicals) by using the method of Spudich and Watt [21] and gel-filtered on 16/60 SephacrylTM S-200 column (GE Healthcare Life Sciences). Purified monomeric actin was stored at 4 ◦C in G-buffer (5 mM Tris/HCl,…