Product Uses Include• Resuspension of actin protein• Dilution and storage of G-actin proteinMaterialThis buffer contains 5 mM Tris-HCl pH 8.0 and 0.2 mM CaCl2. Used as a general G-actin (monomer) buffer with the addition of 0.2 mM ATP (see Cat. # BSA04) and 0.5 mM DTT (A-buffer). The buffer can be changed to a general F-actin (filament supporting) buffer by the addition of 1/10th volume of actin polymerization buffer (see Cat. # BSA02).
For product Datasheets and MSDSs please click on the PDF links below. For additional information, click on the FAQs tab above or contact our Technical Support department at tservice@cytoskeleton.com
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: Can the general actin buffer be stored with ATP added or should it be added fresh?
Answer 1: On the day of the experiment, the general actin buffer (Cat. # BSA01) should be supplemented with 0.2 mM ATP (Cat. # BSA04) to create an optimal actin monomer buffer. We do not recommend storing the actin buffer with ATP in it. Prepare the actin buffer + ATP in a volume that is needed for the experiment. Calcium ions and ATP are required for actin conformation. In addition, ATP is hydrolysed during actin polymerization and is required for the polymerization process.
Question 2: What is the composition of Cytoskeleton’s general actin buffer?
Answer 2: Upon resuspension in 100 ml of sterile de-ionized water, the buffer (Cat. # BSA01) will be at 1X strength buffer and contain: 0.2 mM calcium chloride and 5 mM Tris-HCl, pH 8.0.
If you have any questions concerning this product, please contact our Technical Service department at tservice@cytoskeleton.com
