Binding of ligands to G-actin proves the biological activity of the ligand and identifies the ligand to be a G-actin binding protein. The assays of this kit are based on the spin-down analysis of ligands bound to G-actin. For this purpose G-actin has been conjugated to Sepharose™ beads (ActinBeads) which are ready-to-use.
ActinBeads are pelleted at low centrifugal forces and ligands bound to G-actin will be specifically co-precipitated. SDS-PAGE analysis will reveal binding of G-actin binding ligands. The Trialkit is further used for assays ranging from the determination of binding stoichiometries using gelsolin as reference, up to variation of buffer conditions to possibly optimize G-actin binding.
The kit contains 1x250µl ActinBeads (50% slurry) prepared from rabbit skeletal muscle alpha actin (Cat. # T8020-01) or bovine cardiac alpha actin (Cat. # T8200-01) and buffers to keep actin functional. The Trialkit Manual describes the use of ActinBeads. We guarantee a stable performance of the kit components when stored at -70°C upon arrival for 6 months.