Cell Signaling Technology

Product Pathways - Cytoskeletal Signaling

Phospho-LIMK1 (Thr508)/LIMK2 (Thr505) Antibody #3841

Applications Reactivity Sensitivity MW (kDa) Source
W H (M) (R) Transfected Only 72 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-LIMK1 (Thr508)/LIMK2 (Thr505) Antibody detects transfected levels of LIMK1 and LIMK2 only when phosphorylated at threonine 508 or 505.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr508 of human LIMK1. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from COS cells, untransfected (lane 1), transfected with Wild-type LIMK1 (lanes 2 and 3) or with LIMK1 T508A mutant (lanes 4 and 5), using Phospho-LIMK1 (Thr508)/LIMK2 (Thr505) Antibody (top), LIMK1 Antibody #3842 (middle) or HA-Tag (262K) mAb #2362 (bottom). Cells were either untreated (lanes 1, 2 and 4) or treated with PMA (lanes 3 and 5). (Triple HA-tagged LIMK1 plasmids kindly provided by Dr. K. Mizuno, Biological Institute, Tohoku University, Japan.)

Background

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 (1). LIM kinases are involved in actin cytoskeletal regulation downstream of Rho-family GTPases, PAKs, and ROCK (2,3). PAK1 and ROCK phosphorylate LIMK1 or LIMK2 at the conserved Thr508 or Thr505 residues in the activation loop, increasing LIMK activity (3-5). Activated LIM kinases inhibit the actin depolymerization activity of cofilin by phosphorylation at the amino-terminal Ser3 residue of cofilin (6,7).

  1. Okano, I. et al. (1995) J. Biol. Chem. 270, 31321-31330.
  2. Maekawa, M. et al. (1999) Science 285, 895-898.
  3. Edwards, D. C. et al. (1999) Nat. Cell Biol. 1, 253-259.
  4. Ohashi, K. et al. (2000) J. Biol. Chem. 275, 3577-3582.
  5. Sumi, T. et al. (2001) J. Biol. Chem. 276, 670-676.
  6. Arber, S. et al. (1998) Nature 393, 805-809.
  7. Yang, N. et al. (1998) Nature 393, 809-812.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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