All lanes : Anti-ATP5A antibody (ab129121) at 1/250 dilutionLane 1 : Brain (Human) Tissue Lysate - fetal normal tissue (ab29467)Lane 2 : Heart (Human) Whole Cell Lysate - fetal normal tissueLane 3 : RAW 264.7 (Mouse leukaemic monocyte macrophage cell line) Whole Cell LysateLane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell LysateLane 5 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate Lane 6 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell LysateLane 7 : HL60 (Human promyelocytic leukemia cell line) Whole Cell LysateLane 8 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate Lysates/proteins at 10 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 ATP5A staining in human heart 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 ab129121, 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.