Cell Signaling Technology

Product Pathways - Cytoskeletal Signaling

PAK 1/2/3 Antibody Sampler Kit #4750

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Phospho-PAK1 (Ser144)/PAK2 (Ser141) Antibody #2606 40 µl W H M GP (R) 61 to 67 (PAK2), 68 to 74 (PAK1/3) Rabbit
Phospho-PAK1 (Ser199/204)/PAK2 (Ser192/197) Antibody #2605 40 µl W H M R GP 61 to 67 (PAK2), 68 to 74 (PAK1/3) Rabbit
Phospho-PAK1 (Thr423)/PAK2 (Thr402) Antibody #2601 40 µl W H M GP (R) 61 to 67 (PAK2), 68 to 74 (PAK1/3) Rabbit
Phospho-PAK2 (Ser20) Antibody #2607 40 µl W H M GP (R) 61 to 67 Rabbit
PAK1 Antibody #2602 40 µl W IP IHC-P H M R Mk GP 68 Rabbit
PAK2 (C17A10) Rabbit mAb #2615 40 µl W H M Mk 61 Rabbit IgG
PAK3 Antibody #2609 40 µl W IP H M R 65 Rabbit
PAK1/2/3 Antibody #2604 40 µl W H M R Mk GP 61 (PAK2), 68 (PAK1/3) Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

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 enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Phospho-PAK2 (Ser20) Antibody, PAK1 Antibody, PAK2 (C17A10) Rabbit mAb, PAK3 Antibody and PAK1/2/3 Antibody detect endogenous levels of their target protein(s) and do not cross-react with other PAK family members. Phospho-PAK1 (Ser199/204)/PAK2 (Ser192/197), Phospho-PAK1 (Ser144)/PAK2 (Ser142), and Phospho-PAK1 (Thr423)/PAK2 (Thr402) antibodies, which may also detect Ser200/205, Ser139, and Thr421 of phosphorylated PAK3, respectively. Phospho-PAK1 (Thr423)/PAK2 (Thr402) Antibody cross-reacts with phospho-Mst1 (Thr183) or phospho-Mst2 (Thr180).

Western Blotting

Western Blotting

Western blot analysis of extracts from C2C12 cells, untreated or insulin-treated (100 nM) for 20 minutes, using: Phospho-PAK1 (Thr423)/PAK2 (Thr402) Antibody #2601; PAK1 Antibody #2602; PAK 1/2/3 Antibody #2604; Phospho-PAK1 (Ser199/204)/PAK2 (Ser192/197) Antibody #2605; Phopsho-PAK1 (Ser144)/PAK2 (Ser141) Antibody #2606; Phospho-PAK2 (Ser20) Antibody #2607; PAK3 Antibody #2609; and PAK2 Rabbit mAb #2615.

Description

The PAK antibody sampler kit provides and economical means to evaluate the activation status of PAK1, 2, and 3. This kit includes enough primary and secondary antibodies to perform four western blots with each antibody.

Source / Purification

Phospho-specific antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Ser144, Ser199/204 and Thr423 of human PAK1, and Ser20 of human PAK2. PAK1 Antibody and PAK2 Antibody are produced by immunizing animals with synthetic peptides corresponding to residues surrounding the aminotermini of PAK1 and PAK2. PAK3 Antibody and PAK1/2/3 Antibody is produced by immunizing animals with a synthetic peptide corresponding to residues at the carboxy-terminus of human PAK3 and PAK1. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

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 serines 199 and 204 of PAK1 and serines 192 and 197 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 of Ser144 of PAK1 or Ser139 of PAK3 (located in the kinase inhibitory domain) affects kinase activity (7). Phosphorylation of Ser21 of PAK1 or Ser20 of PAK2 regulates binding with the adaptor protein Nck (8).

  1. Knaus, U.G. and Bokoch, G.M. (1998) Int J Biochem Cell Biol 30, 857-62.
  2. Daniels, R.H. et al. (1998) EMBO J 17, 754-64.
  3. King, C.C. et al. (2000) J Biol Chem 275, 41201-9.
  4. Manser, E. et al. (1997) Mol Cell Biol 17, 1129-43.
  5. Gatti, A. et al. (1999) J Biol Chem 274, 8022-8.
  6. Lei, M. et al. (2000) Cell 102, 387-97.
  7. Chong, C. et al. (2001) J Biol Chem 276, 17347-53.
  8. Zhao, Z.S. et al. (2000) Mol Cell Biol 20, 3906-17.

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!

Protocols

Companion Products

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

Products