Cell Signaling Technology

Product Pathways - Cytoskeletal Signaling

Rho-GTPase Antibody Sampler Kit #9968

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Cdc42 (11A11) Rabbit mAb #2466 40 µl W H M R B (X) 21 Rabbit IgG
Phospho-Rac1/cdc42 (Ser71) Antibody #2461 40 µl W H (M) (R) 28 Rabbit
RhoA (67B9) Rabbit mAb #2117 40 µl W H M R Mk B 21 Rabbit IgG
RhoB Antibody #2098 40 µl W H M R Mk 21 Rabbit
RhoC (D40E4) Rabbit mAb #3430 40 µl W H M Mk (R) 21 Rabbit IgG
Rac1/2/3 Antibody #2465 40 µl W F H M R Mk B (X) 21 Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

Applications Key:  W=Western Blotting  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  X=Xenopus  B=Bovine
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

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 Antibody detects endogenous levels of total Rac1/2/3 proteins. RhoA (67B9) Rabbit mAb recognizes endogenous levels of total RhoA protein. RhoB Antibody recongnizes endogenous levels of RhoB. RhoC (D40E4) XP® Rabbit mAb recongnizes endogenous levels of RhoC.

Western Blotting

Western Blotting

Western blot analysis of extracts from mouse and rat brain tissue using RhoB Antibody #2098.

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 A431 cell extracts, untreated (-) or EGF treated (+), using Phospho-Rac1/cdc42 (Ser71) Antibody #2461 (upper) or Rac1/Cdc42 Antibody #4651 (lower).


Western Blotting

Western Blotting

Western blot analysis of extracts from COS, C6, NIH/3T3 and HEK293 cells, using Rac1/2/3 Antibody #2465.

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 RhoC (D40E4) Rabbit mAb #3430.


Description

The Rho-GTPase Antibody Sampler Kit contains reagents to examine aspects of cell migration, adhesion, proliferation and differentiation in cells. This kit includes enough primary and secondary antibodies to perform four Western blot experiments per each primary antibody.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Thr138 of human Rac1, corresponding to residues surrounding Ser71 of human Rac1/cdc42, and corresponding to residues near the carboxy terminus of RhoB . Polyclonal antibodies are purified by protein A and peptide affinity chromatography.Monoclonal antibody is produced by immunizing animals with synthetic peptides corresponding to residues surrounding Lys135 of human Cdc42, residues near the carboxy terminus of human RhoA, and to residues corresponding to the carboxy terminus of human RhoC.

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.

Application References

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Protocols

Companion Products

Selected rabbit monoclonal antibodies are produced under license (granting certain rights including those under U. S. Patent No. 5,675,063 and/or U.S.S.N. 11/476,277) from Epitomics, Inc.

For Research Use Only. Not For Use In Diagnostic Procedures.

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