Cat. # AR05
Product Uses Include
- In vivo actin polymerization studies (microinjection into muscle cells)
- In vitro motility studies using fluorescent F-actin and muscle myosins
Material
Purified rabbit muscle actin (Cat. # AKL99) has been modified to contain covalently linked rhodamine at random surface lysine residues. An activated ester of rhodamine is used to label the protein. The labeling stoichiometry has been determined to be 0.5 dyes per actin monomer. Rhodamine labeled rabbit muscle actin has an approximate molecular weight of 43 kDa, and is supplied as a pink lyophilized powder. The lyophilized protein is stable for 6 months when stored desiccated to <10% humidity at 4°C. The protein should be reconstituted to 10 mg/ml with distilled water, it will then be in the following buffer: 5 mM Tris-HCl pH 8.0, 0.2 mM CaCl2, 0.2 mM ATP, 5% sucrose, and 1% dextran.
Rhodamine actin from a non-muscle source is also available (Cat. # APHR).
Purity
Protein purity is determined by scanning densitometry of Coomassie Blue stained protein on a 12% polyacrylamide gel. AR05 rhodamine muscle actin was found to be >99% pure (see Figure 1).
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Figure 1. A 100 µg sample of rhodamine muscle actin (molecular weight approx. 43 kDa) was separated by electrophoresis in a 12% SDS-PAGE system, and stained with Coomassie Blue. Protein quantitation was performed with the Precision Red Protein Assay Reagent (Cat. # ADV02). |
Biological activity
The biological activity of rhodamine muscle actin can be determined by its ability to efficiently polymerize into filaments in vitro and separate from unpolymerized components in a spin down assay. Stringent quality control ensures that 90% of the labeled muscle actin can polymerized in this assay, which is similar to the unlabeled product (Cat. # AKL99)
Examples of publications where this product was used
Allen, P. G. (2003). Actin filament uncapping localizes to ruffling lamellae and rocketing vesicles. Nat. Cell Biol. 5, 972-979.
Burakov, A., Nadezhdina, E., Slepchenko, B. and Rodionov, V. (2003). Centrosome positioning in interphase cells. J. Cell Biol. 162, 963-969.
Defacque, H., Egeberg, M., Habermann, A., Diakonova, M., Roy, C., Mangeat, P., Voelter, W., Marriott, G., Pfannstiel, J., Faulstich, H. et al. (2000). Involvement of ezrin/moesin in de novo actin assembly on phagosomal membranes. EMBO J. 19, 199-212.
Loomis, P. A., Zheng, L., Sekerkova, G., Changyaleket, B., Mugnaini, E. and Bartles, J. R. (2003). Espin cross-links cause the elongation of microvillus-type parallel actin bundles in vivo. J. Cell Biol. 163, 1045-1055.
Pelham, R. J., Jr. and Chang, F. (2001). Role of actin polymerization and actin cables in actin-patch movement in Schizosaccharomyces pombe. Nat. Cell Biol. 3, 235-244.
| Product description | Cat. # | Amount | Price & Order |
| Rhodamine actin, skeletal muscle | AR05-B | 10 x 20 µg |
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| AR05-C | 20 x 20 µg |
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| AR05-D | Large quantities |
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