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F-actin visualization kit

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Conventional actins have a relative molecular mass of approximately 43 kDa.  Monomeric actin (G-actin) can self-assemble (polymerize) into microfilaments (F-actin), the fundamental unit of the actin cytoskeleton.  Actins are highly conserved within the eukaryotic kingdom and exist in higher eukaryotes as multigene families.  Isoforms show distinct cellular and sub-cellular expression and localization.  It has been demonstrated that different isoforms have subtly different biochemical properties in vitro which supports functional diversity within isotypes in vivo (1).

The actin cytoskeleton is a highly dynamic structure, a property under the tight regulation of more than 150 actin binding proteins (ABPs) (2, 3).  It is involved in a large number of cellular processes, including muscle contraction, lamellopodia extrusion, cell locomotion, cytokinesis, intracellular transport and cytoplasmic streaming (1).  The morphology of the actin cytoskeleton changes rapidly in response to a wide variety of internal and external stimuli.

Fluorescent phalloidins selectively stain filamentous actin at nanomolar concentrations (6).  They are the reagent of choice for F-actin staining of fixed cells for several reasons:

Kit contents

This kit contains enough reagents for staining 300 coverslips (12 mm circular)
Equipment needed
  1. Fluorescence microscope equipped with filters for rhodamine detection
  2. Microscope slides

Example results

Figure 1 in the gallery shows 3T3 cells under serum starvation conditions (A) and calpeptin stimulated conditions (B) stained with the F-actin Visualization Biochem Kit.  It can be seen that most actin stress fibers disappear under serum starvation conditions, due to the de-activation of the Rho signaling pathway, while calpeptin stimulation of the cells results in a rapid accumulation of actin stress fibers due to the activation of Rho (4)

Product Uses Include

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