RGS·His Antibody, BSA-free (100ug)

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	Use the antibody concentrations recommended by the manufacturer

Protocol tips
Use the antibody concentrations recommended by the manufacturer

Publication protocol

The presence of His6-tagged Epo in COS-7 CM, in CHO CM, and in samples at each step of the purification process was monitored using an anti-His tag antibody. Both standard SDS-PAGE/western blotting (to detect denatured protein) and dot blotting were used. For dot blot analysis, samples were applied directly to nitrocellulose membrane, dried, and fixed by baking at 80ºC. Murine dihydrofolate reductase (DHFR) with a C-terminal His6-tag (vector pQE-16, Qiagene) and human selenium binding protein (hSP56) with an N-terminal His6-tag prepared in our laboratory, both purified on Ni2+-NTA resin, were used as positive controls. Following appropriate blocking, blots were incubated in the presence of one of the following anti-His monoclonal antibodies: anti-polyHistidine (Sigma), penta-His antibody, tetra-His antibody or RSG-His antibody (all from Qiagen), using the antibody concentrations recommended by the manufacturers (Table 1). HRP-conjugated goat anti-mouse IgG (Santa Cruz Biotechnology) was employed as second antibody, and chemiluminescent detection was performed using Super Signal (Pierce).

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Manufacturer protocol

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