Revision 3

#8354Store at -20C

1 Kit

(7 x 20 microliters)

Cell Signaling Technology

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For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Cofilin (D3F9) XP® Rabbit mAb 5175 20 µl 19 kDa Rabbit IgG
Phospho-Cofilin (Ser3) (77G2) Rabbit mAb 3313 20 µl 19 kDa Rabbit IgG
LIMK2 (8C11) Rabbit mAb 3845 20 µl 70 kDa Rabbit IgG
TESK1 (D49D4) Rabbit mAb 4655 20 µl 68 kDa Rabbit IgG
ROCK1 (C8F7) Rabbit mAb 4035 20 µl 160 kDa Rabbit 
Chronophin/PDXP (C85E3) Rabbit mAb 4686 20 µl 31 kDa Rabbit IgG
SSH1 (E1K3W) Rabbit mAb 13578 20 µl 140 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.


The Cofilin Activation Antibody Sampler Kit provides an economical means to evaluate the presence and status of cofilin activation. The kit contains enough primary antibody to perform two western blot experiments per antibody.


Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibodies.


Cofilin and actin-depolymerization factor (ADF) are members of a family of essential conserved small actin-binding proteins that play pivotal roles in cytokinesis, endocytosis, embryonic development, stress response, and tissue regeneration (1). In response to stimuli, cofilin promotes the regeneration of actin filaments by severing preexisting filaments (2). The severing activity of cofilin is inhibited by LIMK or TESK phosphorylation at Ser3 of cofilin (3-5). Phosphorylation at Ser3 also regulates cofilin translocation from the nucleus to the cytoplasm (6).
LIM kinases (LIMK1 and LIMK2) are serine/threonine kinases that have two zinc finger motifs, known as LIM motifs, in their amino-terminal regulatory domains (7). LIM kinases are involved in actin cytoskeletal regulation downstream of Rho-family GTPases, PAKs, and ROCK (8,9). PAK1 and ROCK phosphorylate LIMK1 or LIMK2 at the conserved Thr508 or Thr505 residues in the activation loop, increasing LIMK activity (9-11). Activated LIM kinases inhibit the actin depolymerization activity of cofilin by phosphorylation at the amino-terminal Ser3 residue of cofilin (12,13).
Testis-specific kinase 1 (TESK1) is an LIMK-related protein kinase originally identified to be highly expressed in testes and subsequently shown to be expressed in a wide variety of tissues and cell types (14-17). TESK1 phosphorylates the actin severing protein cofilin at Ser3, inactivating cofilin and thus regulating the organization of the actin cytoskeleton (15). Integrin signaling activates TESK1 activity and leads to stress fiber formation and cell spreading (15,18,19). TESK1 is involved in regulation of ERK signaling through its interaction with Spry2 (20) and regulation of cell spreading through its interaction with the focal adhesion protein actopaxin/α-parvin (18).
Chronophin (CIN, PDXP) is a haloacid dehalogenase phosphatase that dephosphorylates cofilin. Alteration of CIN activity through overexpression of either the wildtype or phosphatase-inactive mutant CIN interferes with actin dynamics, cell morphology and cytokinesis (21).
ROCK (Rho-associated kinase), a family of serine/threonine kinases, is an important downstream target of GTPase Rho and plays an important role in Rho-mediated signaling. Two isoforms of ROCK have been identified (ROCK1 and ROCK2). ROCK is composed of N-terminal catalytic, coiled-coil, and C-terminal PH (pleckstrin homology) domains. The C-terminus of ROCK negatively regulates its kinase activity (22,23). Caspase-3-induced cleavage of ROCK1 and direct cleavage of ROCK2 by granzyme B (grB) activates ROCK and leads to phosphorylation of myosin light chain and inhibition of myosin phosphatase (24). This phosphorylation may account for the mechanism by which Rho regulates cytokinesis, cell motility, cell membrane blebbing during apoptosis, and smooth muscle contraction (25-27).
Slingshot homolog 1 (SSH1) can also dephosphorylate LIMK kinases, suppressing LIMK phosphorylation of cofilin (28). In addition, SSH1 modulates actin dynamics by stabilizing F-actin and promoting actin bundling independent of its cofilin phosphatase activity (29). SSH1 activity is regulated by phosphorylation and protein-protein interaction through various signaling pathways (1). Binding of SSH1 to F-actin stimulates its cofilin phosphatase activity (30).

  1. Carlier, M.F. et al. (1999) J Biol Chem 274, 33827-30.
  2. Condeelis, J. (2001) Trends Cell Biol 11, 288-93.
  3. Arber, S. et al. (1998) Nature 393, 805-9.
  4. Yang, N. et al. (1998) Nature 393, 809-12.
  5. Toshima, J. et al. (2001) J Biol Chem 276, 31449-58.
  6. Nebl, G. et al. (1996) J Biol Chem 271, 26276-80.
  7. Okano, I. et al. (1995) J Biol Chem 270, 31321-30.
  8. Maekawa, M. et al. (1999) Science 285, 895-8.
  9. Edwards, D.C. et al. (1999) Nat Cell Biol 1, 253-9.
  10. Ohashi, K. et al. (2000) J Biol Chem 275, 3577-82.
  11. Sumi, T. et al. (2001) J Biol Chem 276, 670-6.
  12. Arber, S. et al. (1998) Nature 393, 805-9.
  13. Arber, S. et al. (1998) Nature 393, 805-9.
  14. Toshima, J. et al. (1995) J Biol Chem 270, 31331-7.
  15. Toshima, J. et al. (2001) Mol Biol Cell 12, 1131-45.
  16. Toshima, J. et al. (2001) J Biol Chem 276, 31449-58.
  17. Toshima, J. et al. (2001) Biochem Biophys Res Commun 286, 566-73.
  18. LaLonde, D.P. et al. (2005) J Biol Chem 280, 21680-8.
  19. Tsumura, Y. et al. (2005) Biochem J 387, 627-37.
  20. Chandramouli, S. et al. (2008) J Biol Chem 283, 1679-91.
  21. Gohla, A. et al. (2005) Nat Cell Biol 7, 21-9.
  22. Nakagawa, O. et al. (1996) FEBS Lett 392, 189-93.
  23. Lee, J.H. et al. (2004) J Cell Biol 167, 327-37.
  24. Sebbagh, M. et al. (2005) J Exp Med 201, 465-71.
  25. Amano, M. et al. (1996) J Biol Chem 271, 20246-9.
  26. Kureishi, Y. et al. (1997) J Biol Chem 272, 12257-60.
  27. Totsukawa, G. et al. (2000) J Cell Biol 150, 797-806.
  28. Soosairajah, J. et al. (2005) EMBO J 24, 473-86.
  29. Kurita, S. et al. (2007) Genes Cells 12, 663-76.
  30. Kurita, S. et al. (2008) J Biol Chem 283, 32542-52.

Background References

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