All lanes : Anti-DOCK9 antibody (ab70272) at 0.04 µg/ml
Lane 1 : HeLa whole cell lysate at 50 µg Lane 2 : HeLa whole cell lysate at 15 µg Lane 3 : HeLa whole cell lysate at 5 µg
Developed using the ECL technique
Predicted band size : 236 kDa Observed band size : 236 kDa Additional bands at : 117 kDa,200 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 3 minutes
Immunoprecipitation - DOCK9 antibody (ab70272)
HeLa whole cell lysate (1 mg) was immunoprecipitated with ab70272 at 3 µg/mg of lysate (lane 1) or with IgG control (lane 2). Samples were analysed by Western blotting using ab70272 at 0.04 µg/ml and detected by chemiluminescence with a 3 minute exposure time.
ICC/IF image of ab70272 stained HeLa cells. The cells were 4% formaldehyde 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 (ab70272, 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.