ICC/IF image of ab135672 stained MDA-MB-231 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 ab135672 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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.
Anti-ACK1 antibody (ab135672) at 1/50 dilution + K562 cell line lysates at 35 µg
All lanes : Anti-ACK1 antibody (ab135672) at 1/50 dilutionLane 1 : Lysate of non-transfected 293 cellsLane 2 : Lysate of 293 cells transiently transfected with the ACK1 geneLysates/proteins at 2 µg per lane.
Immunohistochemical analysis of formalin fixed and paraffin embedded Human lung carcinoma labelling ACK1 with ab135672 at 1/50 dilution. Antibody was peroxidase-conjugated to the secondary antibody, followed by DAB staining.
Flow cytometric analysis of NCI-H292 cells labelling ACK1 (bottom histogram) with ab135672 at 1/10 dilution and a negative control cell (top histogram). FITC-conjugated goat-anti-rabbit secondary antibodies were used for the analysis.