ab135655 stained A549 cells. The cells were 4% formaldehyde fixed for 10 minutes at room temperature and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab135655 at 5µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
Anti-PTRF antibody - N-terminal (ab135655) at 1/50 dilution + A375 cell line lysate at 35 µg
Immunohistochemistry analysis of formalin-fixed, paraffin-embedded Human lung carcinoma tissue labelling PTRF, with ab135655 at 1/10 dilution followed by peroxidase-conjugated secondary antibody and DAB staining.