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ProteoSep® Buffers

New Buffers for Internal Gradient Chromatofocusing


ProteoSep® 2D Technology incorporates chromatofocusing as a first dimension to separate proteins based on their “pI”. We accomplish this using an internal gradient ran through our ProteoSep® HPCF (weak anion exchange) columns. In the past this was achieved using PolyBuffers (GE Healthcare) to generate this internal gradient but they have since been discontinued.  To address this, Eprogen has developed a replacement for the PolyBuffer 96 and PolyBuffer 74 and has available a pH 9 loading buffer (ProteoSep® SB) and a pH 4 eluent buffer (ProteoSep® EB) which produce linear gradients from a range pH 9- 4 using the ProteoSep® HPCF columns. The formulas are stable and generate the same pH slopes as generated with PolyBuffers.


HPCF is an AX-300 WAX bonded phase that comes in analytical formats [300Å, 6.5um particle sizes] and preparative formats [300Å, 30um particle sizes]. We can produce other particle size materials as well with the same material.

Sample: apo-transferrin

Column: ProteoSep® HPCF (250x2.1 mm)

Flow: 0.2mL/min

Start Buffer: ProteoSep® SB (adjusted with 1N HCl to pH 8.2 )

Elution Buffer: ProteoSep® EB

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