IHC image of Glucocorticoid Receptor (phospho S226) staining in human skeletal muscle 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 ab93104, 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.

All lanes : Anti-Glucocorticoid Receptor (phospho S226) antibody (ab93104) at 1 µg/mlLane 1 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate Lane 2 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate with with immunizing peptide at 1 µg/mlLane 3 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate with with non modified peptide at 1 µg/mlLysates/proteins at 10 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.