Western blot (WB) analysis of DDDDK-tag pAb at 1:5000 dillution
Lane1:HEK293T whole cell lysate
Lane2:HEK293T whole cell lysate,transfected with pcDNA3.1-DDDDK-p62-HA.
IF image of AP0007 stained HEK293T cells, transfected with pcDNA3.1-HA-DDDDK-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 DDDDK-tag pAb #AP0007(1:200) and the antibody HA-tag (4G3) mAb #AP0005M(1:200) at 5µg/ml overnight at +4°C. The secondary antibody (Green) wasGoat anti-Rabbit IgG (H+L) -FITC#BS10950 and the secondary antibody (Red) was Goat anti-Mouse IgG (H+L)-TRITC#BS11502. Hoechst33342 #BD5011 was used to stain the cell nuclei (blue).
Immunoprecipitation of 293F cell transfected with pcDNA3.4-TfR-DDDDK lysates using DDDDK-Tag pAb (Sepharose Bead Conjugate)#BD0048 (lane 2 and lane 3) and Nonspecific IgG Control (Sepharose Bead Conjugate)#BD0048 (lane 4 and lane 5) .Lane 1 is 30% input. The western blot was probed using DDDDK-Tag pAb #AP0007.
