All lanes : Anti-Cdk2 antibody (ab64669) at 1 µg/mlLane 1 : Liver (Human) Tissue Lysate - adult normal tissue (ab29889)Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 3 : Thymus (Rat) Tissue LysateLane 4 : HeLa Whole Cell Lysate - Bleomycin Treated (20U/ml)Lane 5 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell LysateLane 6 : Y79 (Human retinoblastoma cell line) Whole Cell LysateLysates/proteins at 10 µg per lane.SecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilutionPerformed under reducing conditions.
ICC/IF image of ab64669 stained HepG2 cells. The cells were 4% PFA 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 (ab64669, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 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. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa, Hek293, MCF-7 cells at 1µg/ml, and in 100% Methanol fixed (5 min) HeLa, Hek293, HepG2, MCF-7 cells at 5µg/ml.