Revision 1

#4750Store at -20C

1 Kit

(8 x 20 microliters)

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.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Phospho-PAK1 (Ser144)/PAK2 (Ser141) Antibody 2606 20 µl 61 to 67 (PAK2), 68 to 74 (PAK1/3) kDa Rabbit 
Phospho-PAK1 (Ser199/204)/PAK2 (Ser192/197) Antibody 2605 20 µl 61 to 67 (PAK2), 68 to 74 (PAK1/3) kDa Rabbit 
Phospho-PAK1 (Thr423)/PAK2 (Thr402) Antibody 2601 20 µl 61 to 67 (PAK2), 68 to 74 (PAK1/3) kDa Rabbit 
Phospho-PAK2 (Ser20) Antibody 2607 20 µl 61 to 67 kDa Rabbit 
PAK1 Antibody 2602 20 µl 68 kDa Rabbit 
PAK2 (C17A10) Rabbit mAb 2615 20 µl 61 kDa Rabbit IgG
PAK3 Antibody 2609 20 µl 65 kDa Rabbit 
PAK1/2/3 Antibody 2604 20 µl 61 (PAK2), 68 (PAK1/3) kDa Rabbit 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

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 two western blots with each antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

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.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    使用に関する制限

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    Revision 1
    #4750

    PAK 1/2/3 Antibody Sampler Kit

    PAK 1/2/3 Antibody Sampler Kit: Image 1 Expand Image
    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.)
    PAK 1/2/3 Antibody Sampler Kit: Image 2 Expand Image
    Western blot analysis of extracts from SH-SY5Y, C6 and NIH/3T3 cells, using PAK1 Antibody.
    PAK 1/2/3 Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from SH-SY5Y, C6 and NIH/3T3 cells, using PAK1/2/3 Antibody.
    PAK 1/2/3 Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from guinea pig neutrophils stimulated with fMLP (1 µM ) for the indicated times, using Phospho-PAK1 (Ser199/204)/PAK2 (Ser192/197) Antibody.
    PAK 1/2/3 Antibody Sampler Kit: Image 5 Expand Image
    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.
    PAK 1/2/3 Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of extracts from guinea pig neutrophils stimulated with fMLP for the indicated times, using Phospho-PAK2 (Ser20) Antibody (upper) or PAK1/2/3 Antibody #2604 (lower).
    PAK 1/2/3 Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of mouse brain extract and recombinant GST-PAK3, using PAK3 Antibody.
    PAK 1/2/3 Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of extracts from 293, C2C12 and COS cells using PAK2 (C17A10) Rabbit mAb.
    PAK 1/2/3 Antibody Sampler Kit: Image 9 Expand Image
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
    PAK 1/2/3 Antibody Sampler Kit: Image 10 Expand Image
    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).
    PAK 1/2/3 Antibody Sampler Kit: Image 11 Expand Image
    Western blot analysis of extracts from HeLa cells transfected with 100 nM control siRNA #6201 (-) or PAK1 siRNA, using PAK1 Antibody #2602 and p42 MAP Kinase (Erk2) Antibody #9108. The PAK1 Antibody confirms silencing of PAK1 expression, and p42 MAP Kinase (Erk2) Antibody is used to control for loading and specificity of PAK1 siRNA.
    PAK 1/2/3 Antibody Sampler Kit: Image 12 Expand Image
    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).
    PAK 1/2/3 Antibody Sampler Kit: Image 13 Expand Image
    Immunoprecipitation of PAK3 from mouse brain extract, using PAK3 Antibody, followed by Western blot analysis using PAK3 Antibody (left), PAK1 Antibody #2602 (middle) or PAK2 Antibody #2608 (right).
    PAK 1/2/3 Antibody Sampler Kit: Image 14 Expand Image
    Immunoprecipitation of PAK1 from C6 and SH-SY5Y cells followed by Western blot analysis, using PAK1 Antibody.