All lanes : Anti-IL9 antibody (ab134434) at 1 µg/ml (Milk blocking - 3%)Lane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell LysateLane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell LysateLane 3 : THP1 (Human acute monocytic leukemia cell line) Whole Cell LysateLane 4 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell LysateLysates/proteins at 25 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
IHC image of IL9 staining in Human normal lung 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 ab134434, 10µg/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.