ActinRed 555 ReadyProbes Reagent Protocol

ActinRed 555 ReadyProbes Reagent is a selective, high-affinity F-actin probe conjugated to the red-orange fluorescent dye tetramethylrhodamine (TRITC). Like other ReadyProbes reagents, ActinRed 555 reagent is formulated as a room temperature–stable solution and comes in a convenient dropper bottle.

Labeling fixed cells

First, fix and permeabilize cultured cells with a protocol appropriate for your sample. Counterstain as desired. DAPI may be added directly to fixative.

1. Wash the cells 1–3 times in PBS as needed.

2. Add two drops of ActinRed 555 ReadyProbes Reagent per milliliter of medium.

3. Incubate for 30 minutes, protected from light.

4. Remove the stain solution.

5. Wash the cells 2–3 times in PBS.

6. Image the cells.

Multicolor staining of BPAE cells. BPAE cells were fixed and permeabilized using the Image-iT Fixation/Permeabilization Kit. Actin was stained using ActinRed 555 ReadyProbes Reagent, and nuclei were stained with NucBlue Fixed Cell ReadyProbes Reagent.