Cat. # AKL99 & AKL95
Product Uses Include
- Identification and characterization of muscle actin binding proteins
- In vitro actin polymerization studies
- Antibody standard for Western blot analysis
Material
Actin protein has been purified from rabbit skeletal muscle. AKL99 actin is greater than 99% pure and AKL95 is greater than 95% pure. Muscle actin has an approximate molecular weight of 43 kDa. Rabbit muscle actin is supplied as a white lyophilized powder. The lyophilized protein when stored desiccated to <10% humidity at 4°C is stable for 6 months. When reconstituted in distilled water to 10 mg/ml, the protein is in the following buffer: 5 mM Tris-HCl pH 8.0, 0.2 mM CaCl2, 0.2 mM ATP, 5% sucrose, and 1% dextran.
Purity
Protein purity is determined by scanning densitometry of Coomassie Blue stained protein on a 12% polyacrylamide gel. AKL99 consists of >99% pure muscle actin while AKL95 is >95% pure (see Figure 1).
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Figure 1: Figure 1. Purities of rabbit skeletal muscle actin protein. 100 µg of >99% pure (AKL99) and >95% pure (AKL95) rabbit skeletal muscle actin were run on SDS-PAGE gels and stained with coomassie blue. The arrow indicates actin protein, the arrowhead an α-actinin contaminant (115 kDa). The minor impurities in the purified actins are predominantly actin binding proteins such as α-actinin and gelsolin. |
Biological Activity
The biological activity of muscle actinis determined by its ability to efficiently polymerize into filaments (F-actin) in vitro and separate from unpolymerized components in a spin down assay. Stringent quality control ensures that AKL99 produces >90% F-actin and AKL95 produces >80% F-actin in this assay.
Examples of publications where these products were used
Arora, P. D., Glogauer, M., Kapus, A., Kwiatkowski, D. J. and McCulloch, C. A. (2004). Gelsolin mediates collagen phagocytosis through a rac-dependent step. Mol. Biol. Cell 15, 588-599.
Balcer, H. I., Goodman, A. L., Rodal, A. A., Smith, E., Kugler, J., Heuser, J. E. and Goode, B. L. (2003). Coordinated regulation of actin filament turnover by a high-molecular-weight Srv2/CAP complex, cofilin, profilin, and Aip1. Curr. Biol. 13, 2159-2169.
Engqvist-Goldstein, A. E., Warren, R. A., Kessels, M. M., Keen, J. H., Heuser, J. and Drubin, D. G. (2001). The actin-binding protein Hip1R associates with clathrin during early stages of endocytosis and promotes clathrin assembly in vitro. J. Cell Biol. 154, 1209-1223.
Humphries, C. L., Balcer, H. I., D'Agostino, J. L., Winsor, B., Drubin, D. G., Barnes, G., Andrews, B. J. and Goode, B. L. (2002). Direct regulation of Arp2/3 complex activity and function by the actin binding protein coronin. J. Cell Biol. 159, 993-1004.
Ishikawa, T., Cheng, N., Liu, X., Korn, E. D. and Steven, A. C. (2004). Subdomain organization of the Acanthamoeba myosin IC tail from cryo-electron microscopy. Proc. Natl. Acad. Sci. U. S. A. 101, 12189-12194.
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.
McGhie, E. J., Hayward, R. D. and Koronakis, V. (2001). Cooperation between actin-binding proteins of invasive Salmonella: SipA potentiates SipC nucleation and bundling of actin. EMBO J. 20, 2131-2139.
Sagot, I., Rodal, A. A., Moseley, J., Goode, B. L. and Pellman, D. (2002). An actin nucleation mechanism mediated by Bni1 and profilin. Nat. Cell Biol. 4, 626-631.
Upadhyaya, A., Chabot, J. R., Andreeva, A., Samadani, A. and van Oudenaarden, A. (2003). Probing polymerization forces by using actin-propelled lipid vesicles. Proc. Natl. Acad. Sci. U. S. A. 100, 4521-4526.
| Product description | Cat. # | Amount | Price & Order |
| Actin protein, skeletal muscle, >99% pure | AKL99-A | 4 x 250 µg |
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| AKL99-B | 2 x 1 mg |
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| AKL99-C | 5 x 1 mg |
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| AKL99-D | 10 x 1 mg |
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| AKL99-E | 20 x 1 mg |
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| AKL99-XL | Large quantities |
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| Actin protein, skeletal muscle, >95% pure | AKL95-B | 1 x 1 mg |
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| AKL95-C | 5 x 1 mg |
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| AKL95-D | 10 x 1 mg |
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| AKL95-E | 20 x 1 mg |
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| AKL95-XL | Large quantities |
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