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Measurement of Hydrodynamic Radius for a Standard Protein Over a Wide Concentration RangeMay 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. Instrumentation and materials 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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