Skip to main content
. Author manuscript; available in PMC: 2010 Jul 25.
Published in final edited form as: Methods Enzymol. 2000;326:245–254. doi: 10.1016/s0076-6879(00)26058-8

Fig. 2.

Fig. 2

SDS–PAGE analysis of a representative polyhistidine-tagged protein purification using a nickel-nitrilotriacetic acid (Ni2+–NTA) matrix. The 42-kDa MAP-kinase protein ERK2 was affinity tagged at its N terminus with six histidines [D. J. Robbins, E. Zhen, H. Owaki, C. A. Vanderbuilt, D. Ebert, T. D. Geppert, and M. H. Cobb, J. Biol. Chem. 268, 5097 (1993)], and was isolated using the procedure detailed in the text. Shown are equal volumes of the cell lysate, the breakthrough material that failed to bind to the resin during the batch step, the wash material obtained after loading the resin into the column, and the eluate from the column. The 15% Laemmli gel was visualized by Coomassie staining.