Cell Signaling Technology

Product Pathways - Cytoskeletal Signaling

Phospho-PAK1 (Ser144)/PAK2 (Ser141) Antibody #2606

No. Size Price
2606S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
2606T 20 µl ( 2 western blots ) ¥1,500.00 现货查询 购买询价
2606 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Guinea Pig, Endogenous 61 to 67 (PAK2), 68 to 74 (PAK1/3) Rabbit

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting,

Homology

Species predicted to react based on 100% sequence homology: Rat,

Specificity / Sensitivity

Phospho-PAK1 (Ser144)/PAK2 (Ser141) Antibody detects endogenous levels of PAK1 and PAK2 only when phosphorylated at serine 144 or serine 141, respectively. This antibody also recognizes Ser139 phosphorylated PAK3 but does not cross-react with phosphorylated PAK4-6.

Phospho-PAK1 (Ser144)/PAK2 (Ser141) Antibody分别检测内源性PAK1蛋白和serine 144或serine 141位点磷酸化的PAK2蛋白。该抗体也能识别Ser139位点磷酸化的PAK3蛋白,但是不能与磷酸化的PAK4-6发生交叉反应。

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser144 of human PAK1. Antibodies are purified by protein A and peptide affinity chromatography.

通过人工合成人源PAK1蛋白Ser144位点周围相应的多肽片段去免疫动物从而制备出多克隆抗体。通过蛋白A和多肽亲和层析纯化抗体。

Western Blotting

Western Blotting

Western blot analysis of extracts from COS cells cotransfected with activated cdc42 and GST-PAK2 (lane 1), transfected with GST-PAK2 alone (lane 2) or activated cdc42 alone (lane 3), using Phospho-PAK1 (Ser144)/PAK2 (Ser141) Antibody (upper) or PAK1/2/3 Antibody #2604 (lower). This antibody detects GST-PAK2 signal only when it is cotransfected with activated cdc42, indicating that this antibody is phospho-specific.

使用Phospho-PAK1 (Ser144)/PAK2 (Ser141) Antibody (上图)或PAK1/2/3 Antibody #2604 (下图),免疫印迹(Western Blot)分析COS细胞,细胞分别共转染cdc42和GST-PAK2 (第1道)、单转染GST-PAK2(第2道)或单转染活性的cdc42。该抗体检测仅当共转染活性的cdc42的GST-PAK2信号,指出该抗体是磷酸化特异性。

Western Blotting

Western Blotting

Western blot analysis of extracts from guinea pig neutrophils stimulated with fMLP for indicated times, using Phospho-PAK1 (Ser144)/PAK2 (Ser141) Antibody (upper) and PAK1/2/3 Antibody #2604 (lower).

使用Phospho-PAK1 (Ser144)/PAK2 (Ser141) Antibody (上图)和PAK1/2/3 Antibody #2604 (下图),免疫印迹(Western Blot)分析guinea pig neutrophils中Phospho-PAK1 (Ser144)/PAK2 (Ser141)和PAK1/2/3蛋白水平,细胞如图所示时间使用fMLP处理。

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).

丝氨酸/苏氨酸激酶的p21-activated kinase (PAK)家族是从事于多种细胞内过程,包括细胞骨架的重排、MAPK信号、凋亡信号、吞噬细胞NADPH氧化酶的控制和生长因子诱导的轴突生长(1,2)。诱导PAK激活的数种机制已经被报道。Rac/Cdc42结合到PAK的氨基端CRIB (or PBD)结构域上引起PAK的自磷酸和构象的改变(1)。通过 PDK使PAK1在Thr423位点磷酸化诱导PAK1的激活(3)。数个自磷酸化位点已经被鉴定,包括PAK1的Ser199和Ser204位点以及PAK2的Ser192和Ser197位点(4,5)。因为自磷酸化位点位于氨基端抑制结构域,所以推测在这个区域内的修饰抑制该激酶恢复到失活构象(6)。研究指出PAK1的Ser144位点磷酸化或PAK3的Ser139位点(位于该激酶抑制区域)影响激酶活性(7)。PAK1的Ser21位点或PAK2的Ser20位点的磷酸化调节与接头蛋白Nck的结合(8)。PAK4、PAK5和PAK6与PAK1-3在氨基端调节域有低的相同序列(9)。PAK4的Ser474位点的磷酸化类似于PAK1的Thr423位点,它可能在调节PAK4的活性和功能起着重要作用(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.

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Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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