Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human laryangeal squamous cell carcinoma labelling Topoisomerase II alpha with ab12318 at 1/1000 (1µg/ml). Detection: DAB.

IHC image of Topoisomerase II alpha staining in human testis 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 ab12318, 1µg/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.

Detection of Human Topo II Alpha by Western Blot and Immunoprecipitation.

Samples:

Whole cell lysate (25 mcg/lane for A; 0.5 mg for IP in B; 25 mcg for Input in B) from HeLa cells. Antibodies: Affinity purified rabbit anti-Topo II alpha antibody BL983 used at the indicated concentrations for WB (A) and IP (B). Control antibody (Ctrl. in A) and antibody for WB of immunoprecipitated Topo II alpha was a monoclonal antibody to Topo II alpha from another source. Detection: Chemiluminescence with exposure times of 5 minutes (A) or 30 seconds (B).