Phospho-PAK1 (Ser199/204)/PAK2

2605:

Western Blotting

Phospho-PAK1 (Ser199/204)/PAK2 (Ser192/197) Antibody detects endogenous levels of Ser199/204 phosphorylated PAK1 or Ser192/197 phosphorylated PAK2. It may also detect Ser200/205 phosphorylated PAK3, however it does not cross-react with phosphorylated PAK4.

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding serine 199/204 of human PAK1. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western blot analysis of extracts from guinea pig neutrophils stimulated with fMLP (1 µM ) for the indicated times, using Phospho-PAK1 (Ser199/204)/PAK2 (Ser192/197) Antibody.

The p21-activated kinase (PAK) family of serine/threonine kinases is engaged in multiple cellular processes, including cytoskeletal reorganization, MAPK signaling, apoptotic signaling, control of phagocyte NADPH oxidase, and growth factor-induced neurite outgrowth (1,2). Several mechanisms that induce PAK activity have been reported. Binding of Rac/Cdc42 to the CRIB (or PBD) domain near the amino terminus of PAK causes autophosphorylation and conformational changes in PAK (1). Phosphorylation of PAK1 at Thr423 by PDK induces activation of PAK1 (3). Several autophosphorylation sites have been identified, including Ser199 and Ser204 of PAK1 and Ser192 and Ser197 of PAK2 (4,5). Because the autophosphorylation sites are located in the amino-terminal inhibitory domain, it has been hypothesized that modification in this region prevents the kinase from reverting to an inactive conformation (6). Research indicates that phosphorylation at Ser144 of PAK1 or Ser139 of PAK3 (located in the kinase inhibitory domain) affects kinase activity (7). Phosphorylation at Ser21 of PAK1 or Ser20 of PAK2 regulates binding with the adaptor protein Nck (8). PAK4, PAK5, and PAK6 have lower sequence similarity with PAK1-3 in the amino-terminal regulatory region (9). Phosphorylation at Ser474 of PAK4, a site analogous to Thr423 of PAK1, may play a pivotal role in regulating the activity and function of PAK4 (10).

1. Knaus, U.G. and Bokoch, G.M. (1998) Int. J. Biochem. Cell Biol. 30, 857-862.
2. Daniels, R.H. et al. (1998) EMBO J. 17, 754-764.
3. King, C.C. et al. (2000) J. Biol. Chem. 275, 41201-41209.
4. Manser, E. et al. (1997) Mol. Cell. Biol. 17, 1129-1143.
5. Gatti, A. et al. (1999) J. Biol. Chem. 274, 8022-8028.
6. Lei, M. et al. (2000) Cell 102, 387-397.
7. Chong, C. et al. (2001) J. Biol. Chem. 276, 17347-17353.
8. Zhao, Z. et al. (2000) Mol. Cell. Biol. 20, 3906-3917.
9. Abo, A. et al. (1998) EMBO J. 17, 6527-6540.
10. Qu, J. et al. (2001) Mol. Cell. Biol. 21, 3523-3533.

• Kaempchen, K. et al. (2003) Upregulation of the Rac1/JNK signaling pathway in primary human schwannoma cells. Hum. Mol. Genet. 12, 1211-1221. Applications: IC-IF
• Zhan, Q. et al. (2003) p21-Activated Kinase 2 in Neutrophils Can Be Regulated by Phosphorylation at Multiple Sites and by a Variety of Protein Phosphatases. The Journal of Immunology 171, 3785-3793. Applications: Western Blotting
• Cai, D. et al. (2003) The GDP exchange factor AND-34 is expressed in B cells, associates with HEF1, and activates Cdc42. J. Immunol. 170, 969-978. Applications: Western Blotting
• Weber, D. S. et al. (2004) Phosphoinositide-Dependent Kinase 1 and p21-Activated Protein Kinase Mediate Reactive Oxygen Species-Dependent Regulation of Platelet-Derived Growth Factor-Induced Smooth Muscle Cell Migration. Circulation Research 94, 1219-1226. Applications: Western Blotting
• Harfouche, R. et al. (2005) FASEB J 19, 1728-30. Applications: Western Blotting
• Misra, U.K. et al. (2005) J Immunol 175, 2525-33. Applications: Western Blotting
• Nikolic, D.S. et al. (2011) Blood 118, 4841-52. Applications: Western Blotting
• Jung, J. et al. (2011) Oncogene 30, 2264-74. Applications: Western Blotting

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• 2602 PAK1 Antibody
• 2604 PAK1/2/3 Antibody
• 2606 Phospho-PAK1 (Ser144)/PAK2 (Ser141) Antibody
• 2607 Phospho-PAK2 (Ser20) Antibody
• 2608 PAK2 Antibody
• 2609 PAK3 Antibody
• 2461 Phospho-Rac1/cdc42 (Ser71) Antibody
• 2601 Phospho-PAK1 (Thr423)/PAK2 (Thr402) Antibody
• 3241 Phospho-PAK4 (Ser474)/PAK5 (Ser602)/PAK6 (Ser560) Antibody
• 3242 PAK4 Antibody
• 7071 Phototope^® -HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide

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