Alexa Fluor^® 488 Phalloidin allows researchers to fluorescently stain the cytoskeleton through the binding of phalloidin to F-actin. This product is intended for use on fixed and permeabilized samples due to the toxicity associated with phalloidin. After reconstitution the stock solution provides enough material to perform 6000 assays based on a 1:800 dilution and a 100 μl assay volume.

Alexa Fluor^® 488 Fluorescent Properties: Excitation: 495, Emission: 518.

To make a 800X stock, reconstitute the lyophilized material in 750 µl anhydrous DMSO. Fix cells for 15 minutes using fresh, methanol-free 4% formaldehyde, then rinse three times in PBS for 5 minutes each. Once fixed, please follow CST protocol for immunostaining. Following incubation of the primary and secondary antibodies, Alexa Fluor^® 488 Phalloidin can be diluted 1:800 (0.125 μl per 100 μl assay volume) in PBS and added to the cells. Allow to incubate for 15 minutes at room temperature, then rinse once with PBS. Coverslip slides with ProLong^® Gold Antifade Reagent #9071 and examine specimen using appropriate excitation wavelength.

Note: Product usage information for this product has changed effective September 01 2024. Previous recommendations were to use either methanol or DMSO as a solvent for lyophilization of the material; It is now recommended to ONLY use DMSO.