[Babcock_Hall]

doi: 10.1016/s0076-6879(00)26058-8
We are following a paper in which they purified their histidine₆-tagged protein (same protein as ours) using a gradient of imidazole. However, they did not specify the volume of buffers. This affects the steepness of the gradient. What would be a logical choice?  I am inclined to say that 10 column volumes, based on other kinds of chromatography, sounds about right.  EDT: An article in 2000 found in Methods in Enzymology by Barnhorst and Falke suggested a 20-CV gradient.

[Babcock_Hall]

We plan to try a 10-column volume gradient and to see what happens.