Rac and Cdc42 Activator (Cat. # CN02) is useful for efficient activation of Rac1, Rac2, Rac3 and Cdc42 in a variety of cultured cells. The reagent activates Rac and Cdc42 proteins in fibroblasts, neurons, epithelial, endothelial, and hematopoietic cells as well as other primary and immortalized lines. Cells treated with the activator can be subjected to any one of a number of assays that indicate an increase in Rac or Cdc42 activity, including membrane ruffles and lamellipodia (Rac) staining (Cat. # BK005 and Figure 2) and Rac or Cdc42 activity assays by G-LISA™ (Cat. # BK125 and BK127 resp.) See Figure 1 for example of Cdc42 activation measured by the G-LISA assay. EGF is standardized in CN02 by measurement in units, thus 100 ng of EGF is 1 unit of CN02.
Rac/Cdc42 Activator II, epidermal growth factor (EGF) acts through a tyrosine kinase receptor (EGFR) to rapidly stimulate actin reorganization at the cell membrane resulting in membrane ruffling. This primary morphological response, which occurs within 1-10 minutes in Swiss 3T3 cells, has been shown to be mediated through activation of Rac (1). Biochemical assays that quantitate the amount of active (GTP-bound) small G-proteins have demonstrated that both Rac and Cdc42 are rapidly activated (within 30 seconds–2minutes) by EGF stimulation (2, 3). A later, secondary response to EGF treatment is the formation of actin stress fibers which is mediated through the activation of Rho (1). EGF is a useful tool in studying Rho signaling pathways, it should be noted, that EGF activates several other important signal transduction pathways including Ras/Raf/MAPK, JAK/STAT and PI3K/AKT and data should be interpreted accordingly.
