Measurement of Hydrodynamic Radius for a Standard Protein Over a Wide Concentration Range

May 2009

This application note shows use of the ActiPix for determination of hydrodynamic radius of a protein. The method allows proteins to be used in their native form, without any labelling or denaturation. A plug of protein solution is injected into a fused silica capillary, driven through the capillary by application of pressure, and detected using UV area imaging as it passes windows at entrance to and exit from a loop in the capillary. The radius of the protein is determined by analysis of band broadening due to Taylor dispersion. The method is applicable over a wide concentration range and uses only nanolitres of sample.

• ActiPix UV imaging system (Paraytec D100)
• 2 x 200 Multiplex CE cartridge (P/N 131-0020)
• 214 nm wavelength filter (P/N 112-0214)
• Commercial capillary electrophoresis system
• 90 cm 75 x 200 μm fused silica capillary (Polymicro Technologies)
• Albumin from bovine serum (Sigma Aldrich A7906)
• 50 mM borate at pH 8.92
• 500 μL microcentrifuge tubes (Sigma Aldrich 178493)

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