• Monitor actin polymerization in vitro via kinetic fluorescence increase
• Characterization of actin-binding proteins (ABPs)
• Screen for ABP ligands

Kit contents (30 – 100 assays)

• Pyrene-labeled muscle actin
• General actin buffer
• Actin polymerization buffer
• ATP
• Tris-HCl pH 7.5
• 96-well plate

Equipment & materials required

• Protein of interest (POI): purified form, not compatible with cell lysates
• Compound of interest
• Fluorimeter: microplate compatible, Ex:350±20 nm, Em: 407±10 nm, kinetic mode
• Ultracentrifuge: 150,000 g, 4°C, 100 – 500 µl tube volume

The Actin Polymerization Kit tracks actin polymerization through the increased fluorescence of pyrene-labeled actin. When pyrene G-actin (monomeric form) assembles into pyrene F-actin (filamentous form), fluorescence intensifies, allowing polymerization to be monitored over time.

Key characteristics

• Detection method: kinetic fluorescence enhancement
• Signal intensity: 4 – 6-fold enhancement of fluorescent signal from monomer to filament form
• Actin substrate: pyrene-labeled rabbit muscle actin
• Timeframe: Approximately 3 hours