Rabbit polyclonal to phospho NF2 (Merlin) Ser 518 (ab2478) was used at a 1/1000 dilution to detect NF2 by Western blot. Approximately 12 ul of a mouse cardiac myocyte lysate was loaded per lane on a 4-20% Criterion gel for SDS-PAGE. Samples were either mock treated (lane 1) or CLA treated at 4nM, 20 nM or 100 nM (lanes 2,3 and 4 respectively) for 45'. After washing, a 1/5,000 dilution of HRP conjugated Goat-anti-Rabbit IgG (ab7090) was used as secondary antibody.
IHC image of ab2478 staining in human normal colon 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 ab2478, 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.