All lanes : Anti-RBCK1 antibody (ab108479) at 1 µg/ml (Milk blocking - 5%)Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell LysateLane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell LysateLane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell LysateLysates/proteins at 20 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
ICC/IF image of ab108479 stained HepG2 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 ab108479 at 5µg/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.