Actin Technical Tips
Tip 1: Storage and handling of actin
Tip 2: Staining F-actin with phalloidin
For more specific technical tips please view our product pages and datasheets.
Tip #1: Keep the lyophilized actin dry by storing in a desiccator at either 4°C or -70°C. After reconstituting actin as directed on the product's datasheet, experiment-sized aliquots must be snap-frozen to liquid nitrogen before storing at -70°C to preserve actin's activity. To maintain activity, actin should not be frozen at a concentration below 10 mg/ml. When thawing the aliquots, thaw tubes in a room temperature water bath and immediately place on ice until used. Do not re-freeze actin once thawed. Throw away unused thawed actin aliquots.
Tip #2: Polymerized actin (aka filamentous or F-actin) is not stable to freezing, but can be stored at 4°C for 2-4 weeks.
Tip #3: Actin monomers are unstable in the absence of ATP, a divalent cation and dithiothreitol (DTT).
Staining F-actin with phalloidin
When preparing cells for phalloidin staining, remember to fix cells in 4% paraformaldehyde rather than methanol. Methanol disrupts the structure of F-actin and the phalloidin binding site.
