All lanes : Anti-KDM5A / Jarid1A / RBBP2 antibody - ChIP Grade (ab70892) at 0.04 µg/mlLane 1 : HeLa whole cell lysate at 50 µgLane 2 : HeLa whole cell lysate at 15 µgLane 3 : HeLa whole cell lysate at 5 µgLane 4 : 293T whole cell lysate at 50 µgLane 5 : NIH3T3 whole cell lysate at 50 µgdeveloped using the ECL technique
HeLa whole cell lysate (at 1 mg for IP, 20% of IP loaded), with ab70892 at 1µg/ml for WB, after IP at 3 µg/mg lysate. Detection: Chemiluminescence with an exposure time of 10 seconds.
ICC/IF image of ab70892 stained HeLa cells. The cells were 100% methanol fixed (5 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 (ab70892, 5µg/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.