Actin-Toolkit Crystallography
Non-Polymerizing Actin — enzymatically modified rabbit skeletal muscle α-actin
Cat. #: 8050-01
Applications
- Co-crystallization of actin-binding proteins with Non-Polymerizing Actin.
- Stabilization of ligand complexes during crystallization.
- Analysis of conformational changes associated with actin binding.
- Biophysical and biochemical studies requiring monomeric actin under defined conditions.
Description
| Product name | Actin-Toolkit Crystallography |
| Protein | Non-Polymerizing Actin (NP-Actin) |
| Origin | Rabbit skeletal muscle α-actin |
| Molecular mass | 42 kDa |
| Toolkit description |
Non-Polymerizing Actin is enzymatically modified rabbit skeletal muscle α-actin cleaved between Gly42 and Val43. This modification disrupts an important monomer–monomer interaction site and strongly suppresses polymerization under Ca2+-based conditions. NP-Actin remains non-polymerized in the presence of conditions such as 100 mM KCl unless Mg2+ is introduced. Following Mg2+ exchange, polymerization competence can be restored. This conditional behavior makes NP-Actin useful when monomeric actin is required during crystallization or complex formation. The toolkit is designed for co-crystallization of actin-binding proteins with actin and for biophysical or biochemical studies of conformational changes upon complex formation. The ability of NP-Actin to bind actin-interacting proteins remains largely preserved. The kit contains six 1 mg vials of NP-Actin prepared from purified rabbit skeletal muscle actin, G-actin buffer components and Mg-Exchange Buffer. A dedicated handbook describes handling and experimental use. |
| Purity | >99% by scanning densitometry. |
Properties
| Form | Lyophilized, ready for reconstitution. |
| Kit content | 6 × 1.0 mg NP-Actin (lyophilized powder); 1 × 1000 ml G-Actin Buffer with ATP (components supplied as powder); 2 × 1.0 ml Mg-Exchange Buffer; 1 × Toolkit Handbook Crystallography. |
| 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. Reconstitute 1.0 mg NP-Actin with 1.0 ml ultrapure water to obtain 1 mg/ml. |
| Purification notes | Purified from rabbit skeletal muscle; GPC. Non-Polymerizing Actin is subsequently generated by enzymatic cleavage between Gly42 and Val43. |
| Protein concentration | Actin concentration determined by direct UV absorbance at 290 nm; A290 nm, 1% = 0.62. |
| Storage instructions | Store at −70 °C upon arrival. Kit components are stable for 6 months under these conditions. Solubilized 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. |
Further Information
Literature
Crystal structure of polymerization-competent actin.
Klenchin VA, Khaitlina SY, Rayment I. J Mol Biol. 2006;362:140. PMID: 16893553.
