All lanes : Anti-53BP2/ASPP2 antibody (ab70548) at 0.04 µg/mlLane 1 : Whole cell lysate from HeLa cells at 50 µgLane 2 : Whole cell lysate from HeLa cells at 15 µgLane 3 : Whole cell lysate from HeLa cells at 5 µgLane 4 : Whole cell lysate from 293T cells at 50 µgdeveloped using the ECL technique
1mg whole cell lysate from HeLa cells was immunoprecipitated with ab70548 at 3ug/mg of lysate(lane 1) or a control IgG (lane 2). Fr the subsequent blot, 20% of immnunoprecipitate was loaded per lane, and probed with ab70548 at 1ug/ml. Detection: chemiluminescence with exposure time of 30 seconds
IHC image of ab70548 staining in normal human skin formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab70548, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.