All lanes : Anti-RNase H1 antibody (ab56560) at 1 µg/mlLane 1 : Human fibroblasts whole cell lysate - untransfectedLane 2 : Human fibroblasts whole cell lysate - transfected with RnaseH constructLysates/proteins at 25 µg per lane.SecondaryHRP-conjugated sheep anti-mouse IgG polyclonal at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
Overlay histogram showing Jurkat cells stained with ab56560 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab56560, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.