An actin marker for the visualization of F-actin in living cells after plasmid transfection
- Brilliant visualization of F-actin in living cells - perfect imaging of cytoskeletal organization and cellular dynamics
- No interference with cytoskeletal dynamics - unrestricted actin functionality
- Excellent signal-to-noise ratio
- Long-term actin staining with stable cell lines
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|60101||pCMVLifeAct-TagGFP2: plasmid, lyophilized, 20 µg||1|
|60102||pCMVLifeAct-TagRFP: plasmid, lyophilized, 20 µg||1|
|60106||pCAGLifeAct-TagGFP2: plasmid, lyophilized, 20 µg||1|
|60107||pCAGLifeAct-TagRFP: plasmid, lyophilized, 20 µg||1|
LifeAct-Plasmids may be used in many cell types when a transient or stable expression of LifeAct is needed. In this case, researchers can choose between two different promoters, CMV (cytomegalovirus) or CAG (modified chicken β-actin). For many standard cell lines, the CMV promoter is the best choice as it leads to strong expression of LifeAct. However, when working with sensitive cells such as neuronal or embryonic stem cells, it is most often recommended to use the CAG promoter, because it does not have a viral origin and an interference with cellular defense mechanisms (“silencing mechanisms”) is less likely.
After transfection of cells with pLifeAct, F-actin is visualized using the fluorescence markers TagGFP2 or TagRFP**. Stable transfection of cell lines allows long-term actin staining for various applications.
Adenoviral vectors are available for difficult-to-transfect cells or when up to 100 % transgene expressing cells are needed. In this case, it is also possible to choose between the two promoters mentioned before.In addition, different stable cell lines are available, which help facilitate daily lab life. You can immediately start performing experiments without the time-consuming and laborious generation of stable cell lines, while still assuring high-quality standards.
* Original LifeAct publication in Nature Methods:
** Information about TagGFP2 and TagRFP at www.evrogen.com
Dr. Christiane Wiesner
“We use LifeAct-TagRFP and LifeAct-TagGFP2 plasmids in primary human macrophages. The LifeAct probes put us in the position to detect F-actin-rich adhesion structures, without the drawback of disturbing their dynamics. The signal is bright and clear without the background of nonintegrated G-actin. LifeAct gives us the opportunity to highlight F-actin-enriched, cytoskeletal organizations, without the disadvantages of fluorophore-tagged actin overexpression.”
Dr. Christiane Wiesner
University Medical Center