Material and Safety Data Sheet
Fascin (human, recombinant) - 2x50 µg
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Fascin (Fascin 1)
(Actin Binding Protein)
Cat. #: 8411-01
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Applications
Fascin (Fascin 1) is a Ca++-insensitive actin-bundling protein suited for in vitro actin network reconstitution, filament crosslinking/bundling assays, and cell motility/cytoskeleton remodeling studies where controlled actin bundling is required.
Description
| Protein | Fascin (Synonyms: Fascin 1, 55 kDa actin-bundling protein, Singed-like protein, p55) |
| Origin | Human recombinant – untagged. |
| Molecular mass | 55 kDa |
| Protein description | Fascin is a monomeric actin-bundling protein with a molecular mass of 55 kDa. Bundling of actin filaments by fascin forms filopodial bundles that push the membrane forward during cell movement and migration and during cytoskeleton remodeling. Fascin has been shown to be involved in the assembly of microspikes, stress fibers and membrane ruffles. While fascin is absent only from epithelial cells, it is upregulated in multiple forms of human carcinoma. |
| Actin interaction | Ca++-insensitive actin bundling protein. |
Properties
| Form | Lyophilized, ready-to-use. |
| Quantity per unit | 2 × 50 µg |
| Buffer | 150 mM NaCl, 20 mM Hepes pH 7.4, 1 mM DTT and 5% sucrose, when reconstituted with 50 µl ultrapure water to obtain a 1.0 mg/ml solution. |
| Purity & Activity |
Purity: >99% by scanning densitometry from Coomassie G-250 stained SDS-gels.
Bundling activity: 2 µM Atto565-Actin (Cat.#: 8162-01) was incubated with Fascin at a molar ratio of 1:10 and 1:4.1 to verify bundling by fluorescence microscopy. All proteins were used as supplied (no dialysis). The actin stock solution was pre-spun 15 min, 15,000×g, 4°C.
Low speed cosedimentation assay (20,000×g, 30 min) was performed with rabbit skeletal muscle actin (Cat.#: 8101-03) in the presence of sugars. At a molar ratio of 4:1 (actin:fascin), fascin was mixed with F-actin. After 30 min at RT, samples were spun; SDS-PAGE showed fascin was completely pelleted with F-actin.
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| Purification notes | Fascin protein was amplified from human macrophage cDNA and expressed as a GST-tag fusion protein in E. coli. After purification by GST-agarose and subsequent GST-tag cleavage, the pure protein was yielded by gel filtration chromatography. |
| Protein concentration | Determined by OD280 (0.1% = 1.224). |
| Storage instructions | Store at −70°C upon arrival and use within the indicated stability period from date of purchase. The solubilized protein is kept on ice and should be stored at −20°C in glycerol or CryoProtect (Cat.#: 6011-01). Avoid repeated freeze/thaw cycles. |
| Shipping conditions | At ambient temperature. Upon delivery store at −70°C. |
| Remarks | |
| CAS no. |
Buffers and antibodies from Hypermol® are made of the ultrapure reagents in Milli-Q™ water, as described in our publications.
Further Information
Material and Safety Data Sheet
Protein Sequences and Structures on RCSB
References
Reconstitution of contractile actomyosin rings in vesicles.
Litschel T, Kelley CF, Holz D, Adeli Koudehi M, Vogel SK, Burbaum L, Mizuno N, Vavylonis D, Schwille P. Nat Commun. 2021 Apr 15;12(1):2254. doi: 10.1038/s41467-021-22422-7.
Structure, evolutionary conservation, and conformational dynamics of Homo sapiens fascin-1, an F-actin crosslinking protein.
Sedeh RS, Fedorov AA, Fedorov EV, Ono S, Matsumura F, Almo SC, Bathe M. J Mol Biol. (2010) 400:589.
Product DataSheet
Protein Sequence on NCBIProtein Sequences and Structures on RCSB
Material and Safety Data SheetReconstitution of contractile actomyosin rings in vesicles.
Litschel T, Kelley CF, Holz D, Adeli Koudehi M, Vogel SK, Burbaum L, Mizuno N, Vavylonis D, Schwille P.Nat Commun. 2021 Apr 15;12(1):2254. doi: 10.1038/s41467-021-22422-7.
J.Mol.Biol. (2010) 400 p.589
