Revision 1
#2601
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For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:
W
Reactivity:
H M GP
Sensitivity:
Endogenous
MW (kDa):
61 to 67 (PAK2), 68 to 74 (PAK1/3)
Source/Isotype:
Rabbit
UniProt ID:
#Q13153, #Q13177, #O75914
Entrez-Gene Id:
5058, 5062, 5063
Product Usage Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Storage
Specificity/Sensitivity
Phospho-PAK1 (Thr423)/PAK2 (Thr402) Antibody detects endogenous PAK1, PAK2 and PAK3 only when phosphorylated at Thr423, Thr402 and Thr421, respectively. The antibody does not cross-react with phosphorylated PAK4, PAK5 or PAK6. The antibody does cross-react with phospho-Mst1 (Thr183) or phospho-Mst2 (Thr180).
Species predicted to react based on 100% sequence homology
Rat
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr423 of human PAK1. 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 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, PAK5/7, 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). PAK family members are widely expressed, and often overexpressed in human cancer (11,12).
Background References
- 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. et al. (2000) Mol. Cell. Biol. 20, 3906-17.
- Abo, A. et al. (1998) EMBO J. 17, 6527-40.
- Qu, J. et al. (2001) Mol. Cell. Biol. 21, 3523-33.
- Wen, Y.Y. et al. (2014) Expert Opin Ther Targets 18, 807-15.
- Molli, P.R. et al. (2009) Oncogene 28, 2545-55.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
W: Western Blotting
Cross-Reactivity Key
H: Human M: Mouse R: Rat Hm: Hamster Mk: Monkey Vir: Virus Mi: Mink C: Chicken Dm: D. melanogaster X: Xenopus Z: Zebrafish B: Bovine Dg: Dog Pg: Pig Sc: S. cerevisiae Ce: C. elegans Hr: Horse GP: Guinea Pig Rab: Rabbit G: Goat All: All Species Expected
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Revision 1
Western blot analysis of extracts from guinea pig neutrophils stimulated with 1 µM fMLP for indicated times, using Phospho-PAK1 (Thr423)/PAK2 (Thr402) Antibody (Provided by Drs. Qian Zhan and John Badwey, Dept. of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Massachusetts.)
Western blot analysis of extracts from HeLa cells treated with 0.4 M sorbitol for indicted times, using Phospho-PAK1 (Thr423)/PAK2 (Thr402) Antibody (upper) or PAK2 antibody (lower).