Cell Signaling Technology

Product Pathways - Cytoskeletal Signaling

PAK1 Antibody #2602

Applications Reactivity Sensitivity MW (kDa) Source
W IP IHC-P H M R Mk GP Endogenous 68 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  GP=Guinea Pig
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

PAK1 Antibody detects endogenous levels of total PAK1 protein. It does not cross-react with PAK2, PAK3 or other PAK family members.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the amino-terminus of human PAK1. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from SH-SY5Y, C6 and NIH/3T3 cells, using PAK1 Antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells transfected with 100 nM control siRNA #6201 (-) or PAK1 siRNA, using PAK1 Antibody #2602 and p42 MAP Kinase (Erk2) Antibody #9108. The PAK1 Antibody confirms silencing of PAK1 expression, and p42 MAP Kinase (Erk2) Antibody is used to control for loading and specificity of PAK1 siRNA.

IP

IP

Immunoprecipitation of PAK1 from C6 and SH-SY5Y cells followed by Western blot analysis, using PAK1 Antibody.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing cytoplasmic localization, using PAK1 Antibody.

Background

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.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

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

Products