The Actin-Toolkit Fluorescence Microscopy is designed to analyze ligand-induced actin polymerization and the formation of actin filament bundles or networks under defined in vitro conditions.
Rhodamine-Actin can be combined with actin-interacting ligands to visualize effects on G-actin or F-actin by fluorescence microscopy. Directly labeled actin allows actin structures to be visualized without a separate phalloidin-based staining step.
The toolkit additionally contains unlabeled rabbit skeletal muscle α-actin, allowing labeled and unlabeled actin to be mixed when a lower fluorescent fraction is desired.
MonoMix, PolyMix and MgCl2 are included for preparation and polymerization of actin under controlled conditions. A dedicated handbook provides step-by-step experimental setups for fluorescence microscopy with labeled actin.
Purity
>99% by scanning densitometry.
Properties
Form
Lyophilized.
Kit content
4 × 100 µg Rhodamine-Actin (lyophilized powder);
1 × 0.5 mg unlabeled α-skeletal muscle actin (lyophilized powder);
5 × 50 ml MonoMix (lyophilized G-actin ATP buffer);
1 × 1.0 ml PolyMix, 10× stock (lyophilized F-actin ATP buffer, pH 7.4);
1 × 0.5 ml 100 mM MgCl2;
1 × Toolkit Handbook Fluorescence Microscopy;
10 × light-protective tubes and tags.
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.
Native actin purified from rabbit skeletal muscle; final purification by gel permeation chromatography (GPC).
Protein concentration
Actin concentration is determined by direct UV absorbance at A290 nm with correction for fluorophore absorbance where applicable. For unlabeled actin, A290nm 1% = 0.62.
Storage instructions
We guarantee stable performance of the kit components 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.
CAS no.
51005-14-2 (Actin)
Remarks
For research use only. Not for human or veterinary diagnostic or therapeutic applications.