ICC/IF image of ab126573 stained A431 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 (ab126573, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.
All lanes : Anti-Cyclophilin F antibody (ab126573) at 1 µg/mlLane 1 : Human Heart Mitochondrial LysateLane 2 : Heart (Human) Tissue Lysate - adult normal tissue (ab29431)Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell LysateLane 4 : HL60 (Human promyelocytic leukemia cell line) Whole Cell Lysate Lane 5 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 6 : HEK293 (Human embryonic kidney cell line) Whole Cell LysateLysates/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.