Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling Rad9 with ab70810 at 1/1000 (1µg/ml). Detection: DAB.
All lanes : Anti-Rad9 antibody - ChIP Grade (ab70810) at 0.1 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 µgdeveloped using the ECL technique
1mg whole cell lysate from HeLa cells was immunoprecipitated using ab70810 at 3ug/mg of lysate (lane 1) or a control rabbit Ig (lane 2). For the subsequent western blot, 20% of the immunoprecipitate was loaded per lane, and probed with ab70810 at 1ug/ml. Detection: chemiluminescence with exposure time of 30 seconds.
IHC image of ab70810 staining in human normal cervix 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 EDTA (pH9, epitope retrieval solution 2) for 20 mins. The section was then incubated with ab70810, 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.