Revision 1
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB

REACTIVITY:

H M GP

SENSITIVITY:

Endogenous

MW (kDa):

61 to 67 (PAK2), 68 to 74 (PAK1/3)

SOURCE:

Rabbit

UniProt ID:

#Q13153, #Q13177, #O75914

Entrez-Gene Id:

5058, 5062, 5063

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.

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 Reactivity:

Human, Mouse, Guinea Pig

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).

  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. et al. (2000) Mol. Cell. Biol. 20, 3906-17.
  9. Abo, A. et al. (1998) EMBO J. 17, 6527-40.
  10. Qu, J. et al. (2001) Mol. Cell. Biol. 21, 3523-33.
  11. Wen, Y.Y. et al. (2014) Expert Opin Ther Targets 18, 807-15.
  12. 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

WB: 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 All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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Revision 1
#2601

Phospho-PAK1 (Thr423)/PAK2 (Thr402) Antibody

Western Blotting Image 1: Phospho-PAK1 (Thr423)/PAK2 (Thr402) Antibody Expand Image
使用 Phospho-PAK1 (Thr423)/PAK2 (Thr402) Antibody 对已经 1 µM fMLP 刺激规定时间的豚鼠嗜中性粒细胞的提取物进行蛋白质印迹分析(组织由马萨诸塞州哈佛大学医学院生物化学和分子药理学科的 Qian Zhan 和 John Badwey 博士惠赠)。
Western Blotting Image 2: Phospho-PAK1 (Thr423)/PAK2 (Thr402) Antibody Expand Image
使用 Phospho-PAK1 (Thr423)/PAK2 (Thr402) Antibody(上图)或 PAK2 Antibody(下图)对已经 0.4 M 山梨醇处理规定时间的 HeLa 细胞的提取物进行蛋白质印迹分析。