LifeAct-TagGFP2 Protein

Applications:

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

Technical Features:

• Recombinant protein, lyophilized
• C-terminal His-tag
• Established protocols for F-actin staining in living and fixed cells
• Excellent signal-to-noise ratio
• Non-toxic F-actin staining of living samples with superb biocompatibility

Specifications:

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

F-actin Visualization in Living Cells Using LifeAct-TagGFP2 Protein

The LifeAct-TagGFP2 Protein is perfectly suited for the quick and efficient visualization of the cytoskeleton in living cells. For the staining procedure, you can use any method for protein transfer that works for your cells of interest.

Note: Please keep in mind that the actual amount of protein within the cell after protein transfection is generally lower when compared to mRNA or DNA transfection. To adequately visualize intracellular F-actin, you should use a high-quality microscopy equipment.

Live cell imaging of F-actin in Rat1 fibroblasts after LifeAct-TagGFP2 Protein transfer (30 µg/ml, 3 minutes).

Super-Resolution Microscopy (STED) of the Actin Cytoskeleton

Using LifeAct-TagGFP2 Protein, the actin cytoskeleton can be visualized in detail. In this experiment, fixed Rat1 fibroblasts were incubated with LifeAct-TagGFP2 protein in a µ-Slide VI ^0.4, ibiTreat. Simulated emission depletion (STED) microscopy was performed to create a super-resolution image.

Super-resolution microscopy of the actin cytoskeleton in Rat1 fibroblasts using LifeAct-TagGFP2 Protein. Microscopy was performed on the STEDYCON super-resolution STED nanoscopy system (Abberior Instruments GmbH, Göttingen, Germany) with a Plan-Neofluar 100x/1.4 objective lens.

F-actin Visualization in Fixed Cells Using LifeAct-TagGFP2 Protein

In addition to working in living cells, the LifeAct-TagGFP2 Protein is also a fast and easy solution for efficient F-actin staining in a variety of fixed cells.

Note: Methanol and acetone can disrupt actin during the fixation process. Therefore, it is recommended to avoid using any methanol- or acetone-containing fixatives in combination with the LifeAct-TagGFP2 protein.

CHO-K1

HT1080

MCF7

Rat1

Formalin-fixed cells were permeabilized with Triton X-100 and incubated in LifeAct-TagGFP2 Protein solution (10 µg/ml in PBS) for 1 hour.

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)

LifeAct is a registered trademark of ibidi GmbH.

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