Western blot (WB) analysis of His-tag polyclonal antibody at 1:5000 dillution
Lane1:HEK293F whole cell lysate,transfected with pcDNA3.4-Flag-N-RBD-His.
Immunoprecipitation of 293F cell transfected with pcDNA3.4-Flag-N-RBD-His lysates using His-Tag pAb (Sepharose Bead Conjugate)#BD0048 (lane 3 and lane 4) and Nonspecific IgG Control (Sepharose Bead Conjugate)#BD0048 (lane 1 and lane 2) .Lane 5 is 30% input. The western blot was probed using His-Tag pAb #AP0032.
IF image of AP0032 stained HEK293F cells, transfected with pcDNA3.4-Flag-N-RBD-His. 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 His-tag pAb #AP0032(1:200) at 5µg/ml overnight at +4°C. The secondary antibody (Green) was Goat anti-Rabbit IgG (H+L) -FITC#BS10950. Hoechst33342 #BD5011 was used to stain the cell nuclei (blue).