All lanes : Anti-IDH2 antibody (ab94359) at 1 µg/mlLane 1 : Heart (Human) Tissue Lysate - adult normal tissue (ab29431)Lane 2 : Skeletal Muscle (Human) Tissue Lysate - adult normal tissue (ab29330)Lane 3 : Liver (Human) Tissue Lysate - adult normal tissue (ab29889)Lane 4 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466)Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate Lane 6 : DU 145 (Human prostate carcinoma cell line) Whole Cell LysateLane 7 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell LysateLysates/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.
ICC/IF image of ab94359 stained HepG2 cells. The cells were 100% methanol fixed (5 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 (ab94359, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit 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. This antibody also gave a positive result in 100% methanol fixed (5 min) MCF7 cells at 1µg/ml, and in 4% PFA fixed (10 min) HepG2 cells at 1µg/ml.