9 reagents to visualize actin in fixed cells

Cystoskeleton #1 - F-actin visualized with Acti-stain™ 488 (PHDG1) in Swiss 3T3 cells treated with CN03 Rho Activator (DNA staining with DAPI (blue)). Cytoskeleton Inc.

2015 was marked by the increasing number of studies related to cellular cytoskeletal dynamics. In this context, it should be noted that the new generation of live cell-imaging SiR-Actin & SiR-Tubulin stains have been widely embraced by Life Scientists. Launched a few months ago, these fluorescent probes are now regularly cited in the “Material & Methods” section of numerous prestigious publications as indicated by Ali (see his the posts “Live cell imaging tool SiR-actin on JBC frontcover” and “User experience of SiR-Actin and SiR-Tubulin Live Cell Imaging“).

Nevertheless, cytoskeletal interactions are still widely studied in “fixed” cells. In the same manner as for live cell-imaging, these experimental assays require high quality reagents too. With the help of our friends from Cystoskeleton Inc., we have selected 9 premium research tools validated for actin visualization in PFA-fixed cells.

Premium reagents for actin visualization in fixed cells

Acti-stains and Rhodamine Phalloidin bind only to F-actin resulting in low background fluorescence. They are ideal for fine observation of the structure of the cellular cytoskeleton.
We’ve also added to this list 5 popular Small G protein modulators, known to heavily regulate actin. They perfectly complement fluorescent Actin stains to further see cytoskeletal intimate interactions and structures as depicted in the images below.

  1. Acti-stain™ 488 phalloidin (cat. nr PHDG1-A) – Ex/Em Wavelengths: 485/535 FITC filter set
  2. Acti-stain™ 555 phalloidin (cat. nr PHDH1-A) – Ex/Em Wavelengths: 535/585 TRITC filter set
  3. Acti-stain™ 670 phalloidin (cat. nr PHDN1-A) – Ex/Em Wavelengths: 640/680 Cy5 filter set
  4. Rhodamine Phalloidin (cat. nr PHDR1) – a 7aa peptide toxin from Amanita phalloides with high affinity binding capacities to F-actin conjugated with Rhodamine
  5. Rho Activator (cat. nr CN01) – for effective RhoA, RhoB, and RhoC activation.
  6. Rac & Cdc42 Activator (cat. nr CN02) – for potent Rac1, Rac2, Rac3 and Cdc42 activation.
  7. Rho Activator II (cat. nr CN03) – activation of Rho GTPase isoforms by deamidating glutamine-63.
  8. Rho/Rac/Cdc42 Activator I (cat. nr CN04) – Rho GTPase isoforms activation by deamidating glutamine-63 of Rho & glutamine-61 of Rac and Cdc42 in their respective Switch II regions
  9. Cell Permeable Rho Inhibitor (cat. nr CT04) – selective inactivation of GTPases RhoA, RhoB, and RhoC

 

See cytoskeletal details in fixed cells

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Philippe Fixe, PhD
Written by Philippe Fixe, PhD
Philippe Fixe is Marketing Director at tebu-bio, passionate about innovation and R&D in Life sciences, Biotechnology, Medical research, Drug discovery, and also a keen photographer.