Cell Signaling Technology

Product Pathways - Cytoskeletal Signaling

Rho-GTPase Antibody Sampler Kit #9968

Kit Includes Quantity Applications Reactivity MW (kDa) Source
Cdc42 (11A11) Rabbit mAb # 2466 40 microliters W H M R B 21 Rabbit
Phospho-Rac1/cdc42 (Ser71) Antibody # 2461 40 microliters W H (M) 28 Rabbit
Rac1/2/3 (L129) Antibody # 2467 40 microliters W IF-IC H M R Mk B 21 Rabbit
RhoA (67B9) Rabbit mAb # 2117 40 microliters W H M R Mk B 21 Rabbit
Anti-rabbit IgG, HRP-linked Antibody # 7074 100 microliters Goat

Applications Key:  W=Western Blotting  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  B=Bovine

Specificity / Sensitivity

Cdc42 (11A11) Rabbit mAb detects endogenous levels of total Cdc42 protein and does not cross-react with other small GTPases. Phospho-Rac1/cdc42 (Ser71) Antibody detects endogenous Rac1/cdc42 only when phosphorylated at Ser71 and may also recognize phospho-RhoA (Ser73). Rac 1/2/3 (L129) Antibody detects endogenous levels of total Rac1/2/3 proteins. RhoA (67B9) Rabbit mAb recognizes endogenous levels of total RhoA protein, but does not recognize the related proteins RhoB or RhoC.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell types using RhoA (67B9) Rabbit mAb #2117.

Western Blotting

Western Blotting

Western blot analysis of extracts from A431 cells treated with EGF for the indicated times using Phospho-Rac1/cdc42 (Ser71) Antibody #2461 (upper) or Rac1/cdc42 antibody (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell types using Cdc42 (11A11) Rabbit mAb #2466.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell types using Rac1/2/3 (L129) Antibody #2467.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic peptide (KLH-coupled) corresponding to the sequence of human Rac1/2/3 and with a synthetic phosphopeptide (KLH-coupled) corresponding to residues surrounding Ser71 of human Rac1/cdc42. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.Monoclonal antibodies are produced by immunizing rabbits with synthetic peptides (KLH-coupled) corresponding to residues surrounding Lys135 of human Cdc42 and to residues near the carboxy terminus of human RhoA.

Background

The Rho family of small GTPases, including Rho, Rac, and cdc42, act as molecular switches to regulate processes such as cell migration, adhesion, proliferation and differentiation (1). RhoA, RhoB and RhoC are all highly homologous but appear to have divergent biological functions. The best characterized of these proteins, RhoA, regulates acomysin contractility, cytokinesis, focal adhesion assembly and cell polarity (2-5). Mammalian Rac exists as three isoforms (Rac1, Rac2 and Rac3) that show high sequence similarity. Well-characterized Rac1 and cdc42 are ubiquitously expressed and play key signaling roles in cytoskeletal reorganization, membrane trafficking, transcriptional regulation, and cell growth and development (6). Phosphorylation of Rac1 at a putative Akt site (Ser71) may limit Rac1 activity through inhibition of GTP binding (7). Rac2 is expressed in cells of hematopoietic origin, while Rac3 is highly expressed in brain and in many other tissues. The Vav family of guanine-nucleotide exchange factors mediates activation of Rho/Rac family small GTPases (8). Negative regulation of Rho-activity members of the p190 RhoGAP family (p190-A and p190-B) may be controlled by Src phosphorylation of Tyr residues, activating the p190 GAP domain (8-10). Furthermore, Rho GDP dissociation inhibitor (RhoGDI) associates with Rho/Rac to negatively regulate nucleotide exchange membrane localization (11).

  1. DerMardirossian, C. and Bokoch, G.M. (2005) Trends Cell Biol. 15, 356-363.
  2. Bi, D. et al. (2005) Circ. Res. 96, 890-897.
  3. Kimura, K. et al. (2000) J. Biol. Chem. 275, 17233-17236.
  4. Barry, S.T. and Critchley, D.R. (1994) J. Cell Sci. 107 ( Pt 7), 2033-2045.
  5. Van Keymeulen, A. et al. (2006) J. Cell Biol. 174, 437-445.
  6. Wennerberg, K. and Der, C.J. (2004) J. Cell Sci. 117, 1301-1312.
  7. Kwon, T. et al. (2000) J. Biol. Chem. 275, 423-428.
  8. Sordella, R. et al. (2003) Cell 113, 147-158.
  9. Chang, J.H. et al. (1995) J. Cell Biol. 130, 355-368.
  10. Roof, R.W. et al. (1998) Mol. Cell Biol. 18, 7052-7063.
  11. Dovas, A. and Couchman, J.R. (2005) Biochem. J. 390, 1-9.

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