Anti-Pak2

The p21-activated kinase (PAK) family of serine/threonine kinases is engaged in multiple cellular processes, including cytoskeletal reorganization, MAPK signaling, apoptotic signaling, control of phagocyte NADPH oxidase and growth factor-induced neurite outgrowth. Several mechanisms that induce PAK activation have been reported. Binding of Rac/cdc42 to the CRIB (or PBD) domain at the N-terminal region of PAK causes autophosphorylation and conformational change of PAK. Phosphorylation of PAK1 at threonine 423 by PDK induces activation of PAK1. Several autophosphorylation sites have been identified, including serines 199 and 204 of PAK1 and serines 192 and 197 of PAK2. Because the autophosphorylation sites are located in the N-terminal inhibitory domain, it has been hypothesized that modification in this region prevents the kinase from reverting to an inactive conformation. Research indicates that phosphorylation of serine 144 of PAK1 or serine 139 of PAK3, which is located in the kinase inhibitory domain, affects the kinase activity. Phosphorylation of serine 21 of PAK1 or serine 20 of PAK2 regulates its binding with the adaptor protein Nck. More recently identified members PAK4, PAK5 and PAK6 have lower sequence similarity with PAK1-3 in the regulatory N-terminal region. It has been demonstrated that phosphorylation of serine 474 of PAK4, an analogous site to threonine 423 of PAK1, may play a pivotal role in the activity and function of PAK4 kinase. Applications: Suitable for use in Western Blot and Immunoprecipitation. Other applications not tested. Recommended Dilution: Western Blot: 1:1000. Western blot analysis of extracts from HeLa, NIH/3T3, C6,, COS, guinea pig neutrophils (GPN) and HEK293 cells recognizes a 61kD protein, Immunoprecipitation: 1:100, Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.