Actin-Toolkit G-Actin Binding
Bovine cardiac α-actin coupled to Sepharose™ (ActinBeads)
Cat. #: 8200-01
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 | Bovine Cardiac Actin |
| Protein | Cardiac α-actin coupled to Sepharose™ as ActinBeads |
| Origin | Bovine cardiac muscle |
| Molecular mass | 42 kDa |
| Toolkit description |
The Actin-Toolkit G-Actin Binding is designed for functional analysis of ligands that interact with monomeric cardiac actin. Bovine cardiac 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 | Bovine cardiac 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 (bovine cardiac 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 | Purified from bovine cardiac muscle; GPC before immobilization. |
| Protein concentration | Actin concentration is determined before immobilization by direct UV absorbance at A290 nm; A290nm 1% = 0.62. |
| Storage instructions | Store at −70 °C upon arrival. Kit components are stable for 6 months under these conditions. Avoid repeated freeze–thaw cycles. |
| 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. |
| CAS no. | 51005-14-2 (Actin) |
Sepharose is a registered trademark of GE Healthcare.
Further Information
References
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 print version comes with the kit)
Flyer Actin-Toolkits
Material and Safety Data Sheet
Protein Sequence on NCBI