This antibody gave a positive signal in Mouse lung tissue lysate in western blot and MCF7 whole cells in Immunofluorescence.
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ICC/IF image of ab97993 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 ab97993 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. This antibody also gave a positive result in formaldehyde fixed (4%, 10min) HeLa, Hek293,and HepG2 cells at 5ug/ml.
Western blot - Pumilio 1 antibody (ab97993)
Anti-Pumilio 1 antibody (ab97993) at 1 µg/ml + Mouse lung normal tissue lysate - total protein (ab29297) at 10 µg
Secondary Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution developed using the ECL technique