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This is intended as a guide only - optimization may be needed.
Phalloidin is a phallotoxin isolated from Amanita phalloides, the death cap mushroom. Fluorescent phallotoxins stain F-actin at nanomolar concentrations. They are water soluble and produce low non-specific binding.
Dissolve the vial contents in 1.5 mL methanol or DMSO. This will yield a solution with a final concentration of approximately 6 μM.
This stock solution can be further diluted as required for the intended application, we recommend a dilution of approximately 40X.
Stock solutions can be frozen for one year when stored at ≤–20°C, desiccated, and protected from light.
Prepare a 1 mL solution containing 50 to 100 μg/mL lysopalmitoylphosphatidylcholine and 3.7% formaldehyde, add approximately 50 μL of Phalloidin-FITC methanolic stock solution. Apply staining solution to cells and leave for 20 minutes at 4°C. 3X Wash cells with PBS. View coverslip.
Phallotoxins are usually not cell-permeable, but it is possible to label some types of cells. Pinocytosis, microinjections and unknown entry methods by hepatocytes have all been documented. Please conduct a literature search to find an appropriate protocol for your experiments.