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|
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 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.
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.
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).
- Knaus, U.G. and Bokoch, G.M. (1998) Int J Biochem Cell Biol 30, 857-62.
- Daniels, R.H. et al. (1998) EMBO J 17, 754-64.
- King, C.C. et al. (2000) J Biol Chem 275, 41201-9.
- Manser, E. et al. (1997) Mol Cell Biol 17, 1129-43.
- Gatti, A. et al. (1999) J Biol Chem 274, 8022-8.
- Lei, M. et al. (2000) Cell 102, 387-97.
- Chong, C. et al. (2001) J Biol Chem 276, 17347-53.
- Zhao, Z.S. et al. (2000) Mol Cell Biol 20, 3906-17.
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For Research Use Only. Not For Use In Diagnostic Procedures.