Actin-Toolkit G-Actin Binding
Rabbit skeletal muscle α-actin coupled to Sepharose™ (ActinBeads)
Cat. #: 8020-01
Applications
- Verification of biological activity through G-actin binding.
- Functional analysis of ligands binding to G-actin.
- Analysis of pharmacological compounds, proteins or lipids that alter ligand–G-actin interactions.
Description
| Product name | Actin-Toolkit G-Actin Binding |
| Protein | Actin (α-actin), coupled to Sepharose™ as ActinBeads |
| Origin | Rabbit skeletal muscle |
| Molecular mass | 42 kDa |
| Toolkit description |
The Actin-Toolkit G-Actin Binding is designed for functional analysis of ligands that interact with monomeric actin. G-actin is immobilized on Sepharose™ as ActinBeads, providing a solid-phase assay for direct binding studies. Ligands bound to the immobilized G-actin are co-sedimented with the ActinBeads at low centrifugal force and can subsequently be analyzed by SDS-PAGE. Gelsolin is included as a reference G-actin-binding protein. The toolkit can be used to assess ligand binding, compare buffer conditions, investigate changes in ligand–G-actin interaction and determine binding stoichiometries under defined in vitro conditions. The supplied handbook introduces the use of ActinBeads and describes experimental approaches for G-actin binding assays. |
| ActinBeads | Rabbit skeletal muscle G-actin conjugated to Sepharose™; supplied as 50% slurry for low-speed sedimentation assays. |
| Purity | >99% by scanning densitometry before immobilization. |
Properties
| Form | ActinBeads supplied as slurry; additional kit components supplied as indicated. |
| Kit content | 4 × 250 µl ActinBeads (G-actin conjugated to Sepharose™, 50% slurry); 4 × 50 ml MonoMix (lyophilized G-actin ATP buffer); 1 × 0.5 ml 100 mM CaCl2; 1 × 15 µg Gelsolin (mammalian); 5 × syringes and needles; 1 × Toolkit Handbook G-Actin Binding. |
| Actin buffer after reconstitution | 2 mM Tris-Cl, pH 8.2, 0.4 mM ATP, 0.5 mM DTT, 0.1 mM CaCl2 and 0.3% disaccharides when reconstituted with 1.0 ml ultrapure water to obtain a 1 mg/ml actin solution. |
| Related products | MonoMix (Cat. #: 5100-0*) may be used for dilution of G-actin or exchange of ligand buffer into an actin-compatible buffer. |
| Purification notes | Native actin purified from rabbit skeletal muscle; gel permeation chromatography (GPC) before immobilization. |
| Protein concentration | Actin concentration is determined before immobilization by direct UV absorbance at A290 nm; A290nm 1% = 0.62. |
| Storage instructions | Stable performance of the kit components is guaranteed for 6 months when stored at −70 °C upon arrival. Solubilized G-actin should be kept on ice and used within 1 week. Avoid refreezing. |
| Shipping conditions | Shipped at ambient temperature. Upon delivery store at −70 °C. |
| Remarks | For research use only. Not for human or veterinary diagnostic or therapeutic applications. |
Sepharose is a registered trademark of GE Healthcare.
Further Information
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.
Actin stabilizing compounds show specific biological effects due to their binding mode.
Wang S, Crevenna AH, Ugur I, Marion A, Antes I, Kazmaier U, Hoyer M, Lamb DC, Gegenfurtner F, Kliesmete Z, Ziegenhain C, Enard W, Vollmar A, Zahler S. Scientific Reports. 2019;9(1):9731. doi: 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 G-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
