Phospho-Rac1/cdc42 (Ser71) Ant

2461:

Western Blotting

Phospho-Rac1/cdc42 (Ser71) Antibody detects endogenous levels of Rac1/cdc42 only when phosphorylated at serine 71. The antibody may also recognize phospho-RhoA (Ser73).

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser71 of human Rac1/cdc42. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western blot analysis of extracts from A431 cells treated with EGF for the indicated times, using Phospho-Rac1/cdc42 (Ser71) Antibody (upper) or Rac1/cdc42 antibody (lower).

Rac and Cdc42 are members of the Rho-GTPase family. In mammals, Rac exists as three isoforms, Rac1, Rac2 and Rac3, which are highly similar in sequence. Rac1 and Cdc42, the most widely studied of this group, are ubiquitously expressed. Rac2 is expressed in cells of hematopoietic origin, and Rac3, while highly expressed in brain, is also found in many other tissues. Rac and Cdc42 play key signaling roles in cytoskeletal reorganization, membrane trafficking, transcriptional regulation, cell growth and development (1). GTP binding stimulates the activity of Rac/Cdc42, and the hydrolysis of GTP to GDP through the protein's intrinsic GTPase activity, rendering it inactive. GTP hydrolysis is aided by GTPase activating proteins (GAPs), while exchange of GDP for GTP is facilitated by guanine nucleotide exchange factors (GEFs). Another level of regulation is achieved through the binding of RhoGDI, a guanine nucleotide dissociation inhibitor, which retains Rho family GTPases, including Rac and Cdc42, in their inactive GDP-bound state (2,3).

A putative Akt phosphorylation site at Ser71 of Rac1/cdc42 has been identified and confirmed by in vitro kinase assay (4). Phosphorylation at this site may inhibit GTP binding of Rac1, attenuating the signal transduction pathway downstream of Rac1 (4).

1. Wennerberg, K. and Der, C.J. (2004) J Cell Sci 117, 1301-12.
2. Bernards, A. and Settleman, J. (2004) Trends Cell Biol 14, 377-85.
3. Rossman, K.L. et al. (2005) Nat Rev Mol Cell Biol 6, 167-80.
4. Kwon, T. et al. (2000) J. Biol. Chem. 275, 423-428.

• John, G. R. et al. (2004) Interleukin-1beta Induces a Reactive Astroglial Phenotype via Deactivation of the Rho GTPase-Rock Axis. J. Neurosci. 24 (11), 2837-2845. Applications: Western Blotting
• Faraone, D. et al. (2006) Blood 107, 1896-902. Applications: Western Blotting

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