Actin-Toolkit F-Actin Binding (alpha skeletal muscle actin) - 4x1.0 mg
Actin-Toolkit F-Actin Binding
α-actin from rabbit skeletal muscle
Cat. #: 8010-01
..............................................................................................................................................................................................................................................................................................................................................................................................................
Applications
- Proof of the biological activity of a ligand by actin binding;
- Cosedimentation analysis of ligands binding to F-actin;
- Testing of actin filament bundling activity or network formation by ligands;
- Analysis of pharmacological compounds, proteins or lipids altering ligand-actin-interaction;
Description
| Protein | Actin (α-Actin) |
| Origin | Skeletal muscle alpha actin, rabbit |
| Molecular mass | 42 kDa |
| Toolkit description | Binding of ligands to filamentous actin is used as a functional proof to state the biological activity of a ligand and characterizes the ligand to be an actin binding protein. Specific actin binding indicates that the ligand is correctly folded and structurally intact. The Actin-Toolkit F-Actin Binding is a well-established and commonly used method to characterize ligand-actin interaction: at high speed centrifugation ligands binding to individual actin filaments are co-sedimented. At low speed spinning a selective co-sedimentation of actin filament bundles or networks is achieved. Low speed assays instantly characterize ligands to induce actin suprastructures. The Actin-Toolkit F-Actin Binding was developed to characterize the actin-binding properties of ligands (native or recombinant proteins, recombinant fragments etc.) and to support the detection even of cryptic actin-binding sites. Further suitable applications are in vitro compound analysis of the ligand-actin interactions by investigating effects of pharmacological compounds on ligand-actin complexes, as well as the introduction of further ligands (proteins, lipids etc.) to ligand-actin complexes. The kit contains 4×1 mg skeletal muscle actin (Cat. # 8010-03), buffers to keep actin functional and to prevent denaturing. A handbook introduces and guides through the experimental work with actin in F-actin Binding Assays. |
| Purity | >99% by scanning densitometry. |
Properties
| Form | Lyophilized, ready-to-use. |
| Kit content | 4×1 mg Actin (lyophilized powder); 4×50 ml MonoMix (lyophilized G-actin ATP-buffer); 1×1 ml PolyMix, 10× stock (lyophilized F-actin ATP-buffer, pH 7.4); 1×1 ml PolyMix 6, 10× stock (lyophilized F-actin ATP-buffer, pH 6.0); 1×1 ml PolyMix 8, 10× stock (lyophilized F-actin ATP-buffer, pH 8.0); 1×0.5 ml MgCl2 (solution); 1× Toolkit Handbook F-Actin Binding; |
| Buffer | 2 mM Tris-Cl pH 8.2, 0.4 mM ATP, 0.5 mM DTT, 0.1 mM CaCl2, 1 mM NaN3 and 0.3% disaccharides, when reconstituted with 1.0 ml ultrapure water to obtain a 1 mg/ml solution. |
| Related products | For dilution of G-actin or exchange of ligand buffer into actin compatible buffer, MonoMix (Cat. #: 5100-0*) may be used. In the absence of nucleators, the polymerization of G-actin is initiated by PolyMix (Cat. #: 5000-0*). |
| Purification notes | Purified from rabbit skeletal muscle, GPC. |
| Protein concentration | Determined by the Biuret method. |
| Storage instructions | We guarantee stable performance of the kit components for 6 months when stored at -70 °C upon arrival. Solubilized G-actin is kept on ice and should be used within 1 week. Avoid refreezing. |
| Shipping conditions | At ambient temperature. Upon delivery store at -70 °C. |
| Remarks | For Use in Research only. Not for Use in Human or Veterinary Diagnostical or Therapeutical Applications. |
References
Actin stabilizing compounds show specific biological effects due to their binding mode.
Wang, S., Crevenna, A. H., Ugur, I., Marion, A., Antes, I., Kazmaier, U., Hoyer, M., Lamb, D. C., Gegenfurtner, F., Kliesmete, Z., Ziegenhain, C., Enard, W., Vollmar, A., & Zahler, S. (2019). Scientific Reports, 9(1), 9731. https://doi.org/10.1038/s41598-019-46282-w
Bin1 directly remodels actin dynamics through its BAR domain.
Dräger NM, Nachman E, Winterhoff M, Brühmann S, Shah P, Katsinelos T, Boulant S, Teleman AA, Faix J, Jahn TR.
EMBO Rep. 2017 Nov;18(11):2051-2066. doi: 10.15252/embr.201744137. Epub 2017 Sep 11.
Handbook Actin-Toolkit F-actin Binding
(Web version only for demonstration purpose!)
A full print version with protocols come with the kit.
Flyer Actin-Toolkits
Material and Safety Data Sheet
Protein Sequence on NCBI
Actin stabilizing compounds show specific biological effects due to their binding mode.
Wang, S., Crevenna, A. H., Ugur, I., Marion, A., Antes, I., Kazmaier, U., Hoyer, M., Lamb, D. C., Gegenfurtner, F., Kliesmete, Z., Ziegenhain, C., Enard, W., Vollmar, A., & Zahler, S. (2019). Scientific reports, 9(1), 9731. https://doi.org/10.1038/s41598-019-46282-w
Bin1 directly remodels actin dynamics through its BAR domain.
Dräger NM, Nachman E, Winterhoff M, Brühmann S, Shah P, Katsinelos T, Boulant S, Teleman AA, Faix J, Jahn TR.
EMBO Rep. 2017 Nov;18(11):2051-2066. doi: 10.15252/embr.201744137. Epub 2017 Sep 11.
