Product Uses Include
This is a 10 x solution that contains 500 mM KCl, 20 mM MgCl2 and 10 mM ATP in 100 mM Tris, pH 7.5. It is used to add to G-actin solutions in A-buffer (General actin buffer (Cat. # BSA01), 0.2 mM ATP (Cat. # BSA04) and 0.5 mM DTT) to induce polymerization into F-actin.
Xiao et al., 2013. c-Yes regulates cell adhesion at the apical ectoplasmic specialization-blood-testis barrier axis via its effects on protein recruitment and distribution. Am. J. Physiol. Endocrinol. Metab. 304, E145-E159.
Butler et al., 2012. Inhibitory effects of pectenotoxins from marine algae on the polymerization of various actin isoforms. Toxicol. In Vitro. 26, 493-499.
Jiwani et al., 2012. Chlamydia trachomatis Tarp cooperates with the Arp2/3 complex to increase the rate of actin polymerization. Biochem. Biophys. Res. Commun. 420, 816-821.
Fan et al., 2012. A role for γS-crystallin in the organization of actin and fiber cell maturation in the mouse lens. FEBS. J. 279, 2892-2904.
Tsai et al., 2011. 7-Chloro-6-piperidin-1-yl-quinoline-5,8-dione (PT-262), a novel ROCK inhibitor blocks cytoskeleton function and cell migration. Biochem. Pharmacol. 81, 856-865.
Trigili et al., 2011. Mechanism of Action of the Cytotoxic Macrolides Amphidinolide X and J. ChemBioChem.12, 1027-1030.
Takamiya et al., 2005. Overexpression of mutated Cu,Zn-SOD in neuroblastoma cells results in cytoskeletal change. Am. J. Physiol. 288, C253-C259.
Kumar et al., 2004. Functional dissection and molecular characterization of calcium-sensitive actin-capping and actin-depolymerizing sites in villin. J. Biol. Chem. 279, 45036-45046.
Fontao et al., 2001. The interaction of plectin with actin: evidence for cross-linking of actin filaments by dimerization of the actin-binding domain of plectin. J. Cell Sci. 114, 2065-2076.
Zhai et al., 2001. Tyrosine phosphorylation of villin regulates the organization of the actin cytoskeleton. J. Biol. Chem . 276, 36163-36167.
Blader et al., 1999. GCS1, an Arf guanosine triphosphatase-activating protein in Saccharomyces cerevisiae, is required for normal actin cytoskeletal organization in vivo and stimulates actin polymerization in vitro. Mol. Biol. Cell. 10, 581-596.
Question 1: Once resuspended, can left-over actin polymerization buffer be stored at 4°C to maintain activity?
Answer 1: Upon reconstitution (2.0 ml of 100 mM Tris-HCl pH 7.5), the actin polymerization buffer (Cat. # BSA02) will be at 10X strength. Set aside the volume needed for that day’s experiments (This buffer is stable for 30 min on ice) and aliquot the remainder into 100 μl volumes, snap frozen in liquid nitrogen, and stored at -70°C. These stocks are stable for 6 months at -70°C.
Question 2: After resuspension, does the actin polymerization buffer need to be used in a certain amount of time before losing activity?
Answer 2: The actin polymerization buffer (Cat. # BSA02) will be stable for 30 min after thawing the previously frozen aliquots (or using freshly prepared). Keep on ice until ready to use. We do not recommend re-freezing aliquots.
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