Western blot (WB) analysis of over-expressed HA-tagged protein in HEK293T cell lysate, the antibody dilutions are 1:1000 (lane 1) and 1:5000 (lane 2). Each lane was loaded with 10 μg of cell lysate.
IF image of AP0005M stained HEK293T cells, transfected with pcDNA3.1-HA-Flag-p62 #PPL00549-2b. The cells were 4% paraformaldehyde fixed (20 min) and then incubated in 10% normal goat serum for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody HA-tag (4G3) mAb #AP0005M(1:200) and the antibody Flag-tag pAb #AP0007(1:200) at 5µg/ml overnight at +4°C. The secondary antibody (Red) was Goat anti-Mouse IgG (H+L)-TRITC#BS11502 and the secondary antibody (Green) was Goat anti-Rabbit IgG (H+L) -FITC#BS10950 used at a 1/1000 dilution for 1h. Hoechst33342 #BD5011 was used to stain the cell nuclei (blue).
