Phalloidin-TRITC is a fluorescein derivative of Phalloidin, which can specifically label myof lin and display red fluorescence when labeled and can be observed using Tesred channels.In Vitro:1. Preparation of Phalloidin-TRITC working solution
1.1Preparation of the stock solution
Dissolve Phalloidin-TRITC in Methanol to obtain 10 mM of stock solution.
Note: It is recommended to store the stock solution at -20 °C or -80 °C away from light and avoid repetitive freeze-thaw cycles.
1.2 Preparation of Phalloidin-TRITC working solutionDilute the stock solution in serum-free cell culture medium to obtain 1-10 μM of working solution.
Note: Please adjust the concentration of Phalloidin-TRITC working solution according to the actual situation.
2. Cell staining
2.1 Suspension cells (6-well plate)
a.Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time. The cell density is 1×106/mL.
b.Add 1 mL of working solution, and then incubate at room temperature for 30-60 minutes.
c.Centrifuge at 400 g at 4°C for 3-4 minutes and then discard the supernatant.
d.Wash twice with PBS, 5 minutes each time.
e.Resuspend cells with serum-free cell culture medium or PBS.
Observation by fluorescence microscopy or flow cytometry.
2.2 Adherent cells
a.Culture adherent cells on sterile coverslips.
b.Remove the coverslip from the medium and aspirate excess medium.
c.Add 100 μL of working solution, gently shake it to completely cover the cells,and then incubate at room temperature for 30-60 minutes.
d.Wash twice with medium, 5 minutes each time. Observation by fluorescence microscopy.