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 two Western blot experiments per each primary antibody.
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.
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.
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).