All lanes : Anti-GST3 / GST pi antibody (ab117885) at 1 µg/mlLane 1 : Prostate (Human) Whole Cell Lysate - liver cirrhosis (ab30307)Lane 2 : PC3 (Human prostate carcinoma cell line) Whole Cell LysateLane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate Lane 5 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell LysateLane 6 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell LysateLane 7 : Kidney (Human) Tissue Lysate - adult normal tissue (ab30203)Lysates/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.
IHC image of GST3 / GST pi staining in Human normal prostate 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 ab117885, 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.
ab117885 stained MCF-7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab117885 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.