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dollins, I do have an excellent reference for you to read J Chromatogr B Analyt Technol Biomed Life Sci. 2002 Mar 25;769(1):133-44. Strategies for the purification and on-column cleavage of glutathione-S-transferase fusion target proteins.Dian C, Eshaghi S, Urbig T, McSweeney S, Heijbel A, Salbert G, Every bit of the method in the paper is easily customizable and in many cases proven worked in our hands PSP On Thu, Feb 28, 2008 at 5:26 PM, Eric Dollins <[log in to unmask]> wrote: > Dear protein purifiers, > Off topic question: Is there a general rule for how fast you can load, > wash and elute from affinity columns, e.g. glutathione agarose? The > product insert from Sigma says load under gravity flow. For the > volume of cell lysate I have, gravity loading would take an > excruciatingly long time. I want to hook up a peristaltic pump to > speed things along, but don't really have a feel for just how fast one > can load a column in general (I realize this is also dependent on the > construct, the buffer, etc). What about the subsequently wash or > elution? > Thanks for help > Eric > > > -- > D. Eric Dollins, Ph.D. > C266 LSRC, Research Dr. > Duke University Medical Center > Durham, NC 27710 > (919) 681-1668, [log in to unmask] > -- Pius S Padayatti Department of Biochemistry, Structural Biology Division, School of Medicine, RT-500 Case Western Reserve University, Cleveland, Ohio-44106, 216-368-6833