DOCK7 functions as a guanine nucleotide exchange factor (GEF), which activates Rac1 and Rac3 Rho small GTPases by exchanging bound GDP for free GTP. It does not have a GEF activity for CDC42. It is required for STMN1 'Ser-15' phosphorylation during axon formation and consequently for neuronal polarization.
Cell projection, axon. Note=Enriched in the developing axons of hippocampal neurons.
ICC/IF image of ab118790 stained U-87 MG cells. The cells were 4% fomaldeyhde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab118790, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Western blot - Anti-DOCK7 antibody (ab118790)
All lanes : Anti-DOCK7 antibody (ab118790) at 0.1 µg/ml
Lane 1 : HeLa whole cell lysate at 50 µg Lane 2 : HeLa whole cell lysate at 15 µg Lane 3 : 293T whole cell lysate at 50 µg Lane 4 : Jurkat whole cell lysate at 50 µg
Detection of DOCK7 in Immunoprecipitates of HeLa whole cell lysatse (1 mg for IP, 20% of IP loaded) using ab118790 at 6 µg/mg lysate for IP and at 1 µg/ml for subsequent Western blot detection. Detection: Chemiluminescence with an exposure time of 30 seconds.