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Hey Matt, it seems to me that what you're asking for is chromatofocusing. See the official GE documentation: http://www6.gelifesciences.com/aptrix/upp00919.nsf/Content/WD:Chromatofocusin(260949098-R350) The proprietary buffers are a bit expensive, but as you found out, they're a bit complicated to make. I don't know if you'd ruin your Mono Q with a pH gradient. If in doubt, buy one of the dedicated Mono P column. Hope that helps. Andreas Matthew Chu wrote: > Dear All, > > Sorry for off-topic question. Does anyone have any experience in > purifying protein using pH gradient in Mono Q column? > > I have been googling for a whole day, only one paper was found to > mention performing pH gradient in Mono Q, but in a mixture of amine > buffering species, which is a bit too complicated (J. Chromatogr. A 1164 > (2007) 181 - 188. Can Tris-Cl/Tris-base or phosphate buffer give a > linear pH gradient from pH 8.0 to 4.0? Is it usual to perform pH > gradient in Mono Q as I don't want to ruin my Mono Q column... > > Any suggestions are welcome. Thanks in advance! > > Kind regards, > Matt > > > ---------------------------------------------------------------------------- > Matthew LH Chu > PhD Student > School of Pharmacy and Pharmaceutical Sciences > University of Manchester > ----------------------------------------------------------------------------