ActinRed™ 555 ReadyProbes® Reagent Protocol
F-actin stain for fixed cells
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
This protocol can be used for:
- F-actin staining in fluorescence microscopy
This protocol should not be used for:
- Flow cytometry
You will need the following for this protocol:
- Cells growing in culture
- ActinRed™ 555 ReadyProbes® Reagent (Cat. No. R37112)
- Fluorescence microscope
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 ActinGreen™ 488 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.|
|Standard filter set||TRITC|
|EVOS® Light Cube||TRITC|
|Storage conditions||Room temperature|
- In some cases, more or fewer drops may be needed to achieve optimal staining intensity.
- Image quality may be improved by replacing the culture medium with Live Cell Imaging Solution (Cat. No. A14291DJ).
For Research Use Only. Not for use in diagnostic procedures.