pCMV/pCAG-LifeAct Plasmids

Applications:

• Cytoskeleton organization:
  - Non-toxic actin visualization
  - Fluorescence staining
• Cytoskeletal dynamics:
  - Actin dynamics in cellular processes
  - Live cell imaging and microscopy

Technical Features:

• For transient and stable transfections
• Superb biocompatibility - non-toxic staining of living samples

Specifications:

Watch our Movie:

Live Cell Imaging of Actin Dynamics in Primary Dendritic Mouse Cells, Transfected with p^CMVLifeAct-TagGFP2

LifeAct, a 17-amino acid peptide, is derived from a protein from Saccharomyces cerevisiae. LifeAct stains filamentous actin (F-actin) structures in living or fixed eukaryotic cells and tissues. In contrast to GFP-actin and its alternatives such as actin-binding proteins, LifeAct does not interfere with actin dynamics in vitro and in vivo, as shown in several cell types and model organisms.*

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.

Method Comparison: Using LifeAct for F-Actin Visualization in Living Cells

¹ Not all of these combinations have been tested in all indicated cell types. We encourage you to use this table as a guideline to understand how the LifeAct system can be optimally applied.

* Original LifeAct publication in Nature Methods:
Riedl J, et al. Lifeact – a versatile marker for the visualization of F-actin. Nature Methods 5, 605-607 (2008)

** Information about TagGFP2 and TagRFP at www.evrogen.com

LifeAct is a registered trademark of ibidi GmbH.

For license and product use information please refer to www.ibidi.com, Legal Notes.