Western blot analysis of E. coli lysate containing 6-His-tagged protein with anti-6-His-tag antibody. Western blot analysis was performed on 160 ng, 80 ng or 20 ng of E. coli lysate containing 6-His-tagged protein. Cells were lysed with SDS sample buffer. The PVDF membrane was blocked with 5% dry milk in PBST for 1 h at RT. The primary antibody anti-6-His-tag mouse monoclonal, 6His (Cat. No. 910HISL) was diluted in blocking buffer (antibody concentration 0.125 μg/ml) and incubated for 1 h at RT. The secondary antibody goat anti-mouse IgG polyclonal, HRP conjugate was also diluted in blocking buffer (antibody concentration 0.2 μg/ml) and incubated for 1 h at RT. The bands were visualized by chemiluminescent detection using Pierce™ ECL Western Blotting Substrate.

Western blot analysis of E. coli lysate containing 6-His-tagged protein with anti-6-His-tag antibody. Western blot analysis was performed on 50 ng or 25 ng of E. coli lysate containing 6-His-tagged protein. Cells were lysed with SDS sample buffer. The PVDF membrane was blocked with 5% dry milk in PBST for 1 h at RT. The primary antibody anti-6-His-tag mouse monoclonal, 6His (Cat. No. 910HISL) was diluted in blocking buffer (antibody concentration 0.05 μg/ml) and incubated for 1 h at RT. The secondary antibody goat anti-mouse IgG polyclonal, HRP conjugate was also diluted in blocking buffer (antibody concentration 0.2 μg/ml) and incubated for 1 h at RT. The bands were visualized by chemiluminescent detection using Pierce™ ECL Western Blotting Substrate.