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Cofilin (Human Recombinant)

Cofilin (Human Recombinant)

Item no.	CF01-A
Manufacturer	Cytoskeleton
Amount	1 x 100 ug
Quantity options	1 x 100 ug 4 x 100 ug 1 x 1 mg
Category	Peptides & Proteins Recombinant Proteins Cell Biology Cytoskeleton Microfilaments
Type	Proteins Recombinant
Specific against	Human (Homo sapiens)
Purity	Protein purity is determined by scanning densitometry of Coomassie Blue stained protein on a 4-20% gradient polyacrylamide gel. The Cofilin protein is 95% pure (see Figure 1).
Citations	van der Gucht, J., Paluch, E., Plastino, J. and Sykes, C. (2005). Stress release drives symmetry breaking for actin-based movement. Proc. Natl. Acad. Sci. U. S. A. 102, 7847-7852. 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. Vignjevic, D., Yarar, D., Welch, M. D., Peloquin, J., Svitkina, T. and Borisy, G. G. (2003). Formation of filopodia-like bundles in vitro from a dendritic network. J. Cell Biol. 160, 951-962. Yokoo, T., Toyoshima, H., Miura, M., Wang, Y., Iida, K. T., Suzuki, H., Sone, H., Shimano, H., Gotoda, T., Nishimori, S. et al. (2003). p57Kip2 regulates actin dynamics by binding and translocating LIM-kinase 1 to the nucleus. J. Biol. Chem. 278, 52919-52923. Idrissi, F. Z., Wolf, B. L. and Geli, M. I. (2002). Cofilin, but not profilin, is required for myosin-I-induced actin polymerization and the endocytic uptake in yeast. Mol. Biol. Cell 13, 4074-4087.
ECLASS 10.1	32160409
ECLASS 11.0	32160409
UNSPSC	12352202
Alias	Actin, Actin Binding Protein, cofilin protein, cofilin, cofilin isotype 1
Similar products	Actin, cofilin, Actin Binding Protein, cofilin protein, cofilin isotype 1
Shipping condition	Room temperature
Available
Shipping Temperature	AT
Storage Conditions	On Arrival: 4°C
Delivery Time	1-2 Weeks
FAQs	Question 1: After reconstituting the lyophilized protein with water, what is the composition of the buffer the cofilin protein is in? Answer 1: The protein should be reconstituted to 5 mg/ml by the addition of 20 µ, l of distilled water. The protein will be in the following buffer, 10 mM Tris pH 8.0, 1 mM EGTA pH 8.0, 5% sucrose, and 1% dextran. In order to maintain high biological activity of the protein, it is strongly recommended that the protein solution be supplemented with DTT to 1 mM final concentration Question 2: To separate the F-actin bound to cofilin, is it absolutely necessary to spin the sample at 100, 000 x g? Answer 2: If possible we recommend spinning the F-actin/cofilin complex at 100, 000 x g to absolutely insure that the protein complex pellets. Check with your university&rsquo, s core facilities or neighboring labs for access to an ultracentrifuge. However, in a pinch, spinning at 50, 000 x g should pellet the majority of the complex.
Weight (grams)	7
Product Uses	Studies of cofilin binding and severing activities Control protein for actin binding protein studies
Material	The human cofilin protein (isotype 1) has been produced in a bacterial expression system. The recombinant protein has a molecular weight of approximately 21 kDa, and does not contain a protein purification tag. Recombinant cofilin has been purified by ion exchange chromatography. Cofilin is one member of a large group of proteins characterized as "actin binding proteins" (ABPs). Cofilin is an essential cellular protein that can bind the barbed end of actin. In the cell, cofilin acts in concert with other regulatory proteins to mediate the response of the actin cytoskeleton to extracellular signals. In vertebrates, cofilin is regulated by pH, phosphorylation and phosphoinositides. Recombinant cofilin is supplied as a white lyophilized powder. The lyophilized protein is stable at 4C desiccated (<10% humidity) for 1 year.When reconstituted with nanopure water, the protein will be in the following buffer: 10 mM Tris pH 8.0, 1 mM EGTA, 5% sucrose and 1% dextran. The reconstituted protein can be stored at -70C for up to 6 months or at 4C for up to 2 weeks with the addition of 100 ug/ml ampicillin and 5 ug/ml chloramphenicol witout any noticeable loss of activity.
Biological Activity	The biological activity of recombinant cofilin is determined by its ability to bind and sever F-actin in a pH dependent manner. Below pH 7.0 cofilin binds to F-actin in a 1:1 molar ratio of cofilin to actin monomer in the filament. Above pH 7.0 cofilin will sever actin filaments and bind actin monomer in a 1:1 molar ratio. A standard biological assay for monitoring the actin binding and severing activity of cofilin consists of SDS-PAGE analysis of F-actin/cofilin spin down assays performed at pH 6.8 and 8.0. Stringent quality control ensures that at pH 6.8 only 20% of cofilin and actin are found in the supernatant, and that a 1:1 molar ratio of cofilin to actin protein is present in the pellet. Furthermore, at pH 8.0 approximately 80% of cofilin and actin are found in the supernatant due to the F-actin severing activity of cofilin.
Figure 1 Legend	Cofilin protein purity determination. A 20 ug sample of CF01 (cofilin molecular weight approx. 21 kDa) was separated by electrophoresis in a 4-20% SDS-PAGE system, and stained with Coomassie Blue.

Note: The presented information and documents (Manual, Product Datasheet, Safety Datasheet and Certificate of Analysis) correspond to our latest update and should serve for orientational purpose only. We do not guarantee the topicality. We would kindly ask you to make a request for specific requirements, if necessary.

All products are intended for research use only (RUO). Not for human, veterinary or therapeutic use.

CF01-A.pdf

CF01-A-datasheet.pdf

CF01-A-safety-datasheet.pdf