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LifeAct-TagGFP2 Protein

New LifeAct-TagGFP2 Protein View larger

A recombinant protein for fast staining and immediate analysis of F-actin in living and fixed cells

  • Brilliant visualization of F-actin for the investigation of cytoskeletal organization and cellular dynamics
  • Fastest staining of F-actin in living cells—get your results within minutes
  • Non-toxic alternative to phalloidin for the staining of living and fixed cells
  • Unrestricted actin functionality—no interference with cytoskeletal dynamics

More details

Product Details

Applications:

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:

Form Lyophilized
Storage -80 °C
TagGFP2 (Exmax / Emmax) 483 / 506 nm


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 the 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, we recommend using Fuse-It-P, ibidi’s reagent for efficient protein transfection using membrane fusion.

Phase Contrast

LifeAct-TagGFP2

Infrared Control Dye

CHO-K1 cells were fused for 5 minutes with Fuse-It-P containing LifeAct-TagGFP2 Protein immediately before microscopy.

Phase Contrast

LifeAct-TagGFP2

Infrared Control Dye

Embryonic rat myofibroblasts were fused for 10 minutes with Fuse-It-P containing LifeAct-TagGFP2 Protein and immediately imaged.

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.

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.


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.

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

Method Suitable for 1
Cell lines
+++
+++
+++
+++
Primary cells
+
++
+++
+++
Non-mammalian cells
-
+
-
-
Transfection Stability
Transient
Yes
Yes
Yes
No
Stable
Yes
No
No
Yes
Protocol and Analysis
Protocol simplicity
+++
++
+++
+++
LifeAct readout start
1–2 days
few minutes
1–2 days
1–2 days
LIfeAct readout duration
several days
(infinity if stable)
up to 1 day
several days
several days
(infinity if stable)
Signal intensity
+++
+
+++
+++
Extra Benefits
Prevention of overexpression artefacts
-
+++
+
+
Prevention of viral response
+++
+++
-
-
Biocompatibility of transfer method
+
+++
++
++
Actin functionality
+++
+++
+++
+++
Safety Consideration
Biosafety Level
BSL1
-
BSL2
BSL2


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.

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