Western blot (WB) analysis of DDDDK-tag (1A8) mAb at 1:10000/1:20000/1:30000 dillution
Lane1:HEK293T whole cell lysate ,untransfected
Lane2:HEK293T whole cell lysate , transfected with pcDNA3.1-HA-DDDDK-p62
Western blot (WB) analysis of DDDDK-tag (1A8) mAb at 1:5000 dilution
Lane1:HEK293T whole cell lysate ,untransfected
Lane2:HEK293T whole cell lysate , transfected with pcDNA3.1-HA-DDDDK-p62 (Anti-SQSTM1, IP)
Lane3:HEK293T whole cell lysate , transfected with pcDNA3.1-HA-DDDDK-p62 (Anti-DDDDK, IP)
Lane4:HEK293T whole cell lysate , transfected with pcDNA3.1-HA-DDDDK-p62 (Anti-DDDDK, IP, supernatant)
IF image of AP0007M stained HEK293T cells,transfected with pcDNA3.1-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 (1A8) #AP0007M(1:200) at 5µg/ml overnight at +4°C. The secondary antibody (Green) was Goat Anti-Mouse IgG (H+L) FITC#BS50950 used at a 1/1000 dilution for 1h. Hoechst33342 #BD5011 was used to stain the cell nuclei (blue).