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Before staining actin with phalloidin dye conjugates, how should I fixate my sample?
Posted January 18, 2018

For example, should methanol be used to fix cell samples prior to staining with Alexa Fluor®-Phalloidin, iFluor-Phalloidin or any other phalloidin conjugates such as FITC or TAMRA?


Answer

Avoid using fixatives that contain methanol because it can disrupt actin during the fixation process. The preferred fixative is methanol-free formaldehyde. You should perform formaldehyde fixation with 4.0% formaldehyde in PBS at room temperature for 10-30 minutes.