Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma tissue labelling MDC1 with ab11170 at 1/1000 (1µg/ml). Detection: DAB.

IHC image of MDC1 staining in human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond 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 ab11170, 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.

Samples:

A) Nuclear extract (50 mcg) from HeLa cells. B) Whole cell lysate from HEK 293 cells. Antibodies: A) ab11170 used at the indicated concentrations for WB. B) ab11170 (column labelled BL579) and ab11171 (column lablled BL580) used at 3.3 mcg/mg lysate for IP followed by WB using ab11169 (column lablled BL578) at 0.1 mcg/ml. Detection: Chemiluminescence with exposure times less than 5 min. Samples: A) Nuclear extract (50 mcg) from HeLa cells. B) Whole cell lysate from HEK 293 cells. Antibodies: A) ab11170 used at the indicated concentrations for WB. B) ab11170 (column labelled BL579) and ab11171 (column lablled BL580) used at 3.3 mcg/mg lysate for IP followed by WB using ab11169 (column lablled BL578) at 0.1 mcg/ml. Detection: Chemiluminescence with exposure times less than 5 min.