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Technical Data Sheet

CK6040

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Cofilin Phospho-Regulation
Antibody Sampler Kit
Price

$395

Kit Components
Applications: WB = Western blot, E = ELISA, ICC = Immunocytochemistry, IP = Immunoprecipitation, IHC = Immunohistochemistry Species: H = Human, R = Rat, M = Mouse, C = Chicken, F = Fish

Product References
SP1711 Wang, Y. et al. (2015) Cancer Lett. 360(2):171
CP1131 Giacobbe, A. et al. (2015) Oncogene. doi: 10.1038 WB: MCF-7
CP1151 Giacobbe, A. et al. (2015) Oncogene. doi: 10.1038 WB: MCF-7
SP1711 DiPaolo, B.C. et al. (2013) Am Jou of Phys 305(2): L141. WB: rat alveolar type II
SP1711 Wang, Y. et al. (2013) J Neurosci. 33(15):6423. WB: Rat infralimbic cortex
SP1711 Sparrow, N. et al. (2012) J Neurosci. 32(15):5284. WB & ICC: Rat Schwann cells
CP1131 Amelio, I. et al. (2012) J Cell Biol. 199(2):347. WB: human primary epidermal keratinocytes
CP1131 Ghia, E.M. et al. (2011) J Immunology. 186:6338. WB: human U937 cells
CP1131 Couch, B.A. et al. (2010) J Alzheimer’s Disease. 20:1003. WB: mouse hippocampus, fasudil
CP1131 Garg, P. et al. (2010) J Biol Chem. 285:22676. WB: human, mouse podocytes
CP1151 Couch, B.A. et al. (2010) J Alzheimer’s Disease. 20:1003. WB: mouse hippocampus, fasudil
LP1891 Marshall, T.W. et al. (2009) J Cell Sci. 122:3061. WB: rat MTLn3 cells
SP1711 Eiseler, T. et al. (2009) Nat Cell Biol. 11(5):545. WB, IP, ICC: human HeLa cells
SP1711 Kim, J.S. et al. (2009) Mol Biol Cell. 20(11):2650. WB: human HeLa, siRNA
SP1711 Yuen, E.Y. & Yan, Z. (2009) J Neurosci. 29:550. ICC: rat interneurons, siRNA
CP1151 Rufanova, V.A. et al. (2009) Cell Health and Cytoskel. 1:17. WB: Glomerular mesangial cell
Kit Summary
The cofilin phospho-regulation antibody sampler kit can be used to monitor the level of phosphorylated cofilin relative to total cofilin expression. In addition, the kit provides reagents for examining the activity and expression levels of the cofilin kinase, LIMK1, and cofilin phosphatase, Slingshot.
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Western blot image of activated mouse recombinant LIMK1 untreated (lanes 1 & 3) or treated with lambda phosphatase (lanes 2 & 4). The blots were probed with anti-LIMK1 (C-terminus) (lanes 1 & 2; LP1831) and anti-LIMK1 (Thr-508) (lanes 3 & 4; LP1891).

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Immunocytochemical labeling in chick dorsal root ganglion neurons using anti-Cofilin (N-terminus; CP1131), anti-Cofilin (Ser-3; CP1151), anti-βIII-Tubulin (C-terminus; TP1691) and anti-β-Tubulin (TM1541) antibodies. (Images provided by Dr. Diane Snow, Department of Anatomy & Neurobiology, University of Kentucky).

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Background
Members of the actin depolymerizing factor (ADF)/cofilin family, cofilin 1, cofilin 2, and ADF are actin-severing proteins that regulate actin remodeling during cellular events such as cell migration, cytokinesis, phagocytosis, endocytosis, axon development, and immune cell activation. Regulation of cofilin activity can occur through phosphorylation of Ser-3 by LIMK1 or LIMK2. LIM kinases are involved in actin cytoskeletal regulation downstream of Rho-family GTPases, PAKs, and ROCK. PAK1 and ROCK phosphorylate LIMK1 or LIMK2 in the activation loop at Thr-508 or Thr-505 residues, respectively. This increases LIMK activity leading to phosphorylation of cofilin at Ser-3. This phosphorylation disrupts cofilin binding to actin in vitro and in vivo. In contrast, Slingshot can dephosphorylate Ser-3 in cofilin promoting actin binding. In mammals, the Slingshot family includes SSH1L, SSH2L, and SSH3L. SSH1L and SSH2L mRNAs are widely expressed, while SSH3L has high expression in epithelial tissues. SSH1L can associate with F-actin and may be the major phosphatase regulating cofilin activity.

Background References
Okano, I. et al. (1995) J. Biol. Chem. 270:31321.
Edwards, D. C. et al. (1999) Nat. Cell Biol. 1:253.
Bamburg, J.R. (1999). Annu Rev Cell Dev Biol. 15:185.
Aizawa, H., et al. (2001). Nature Neurosci. 4(4):367.
Niwa, R. et al. (2002) Cell 108:233.
Maciver, S.K. & Hussey, P.J. (2002). Genome Biol. 3(5):3007.
Endo, M. et al. (2003) J Neurosci. 23(7):2527.
Soosairajah, J. et al. (2005) EMBO J. 24(3):473.
Kobayashi, M. et al. (2006) EMBOJ 25:713.
Buffer and Storage
Rabbit polyclonal antibodies are supplied in phosphate-buffered saline (PBS), 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
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