Conductivity properties of carrier ampholyte pH gradients in isoelectric focusing |
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Authors: | Stoyanov Alexander V Das Champak Fredrickson Carl K Fan Z Hugh |
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Institution: | Department of Mechanical and Aerospace Engineering, McKnight Brain Institute, University of Florida, Gainesville 32611, USA. |
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Abstract: | The conductivity properties of natural pH gradient created by carrier ampholytes were studied during the process of isoelectric focusing (IEF). IEF was performed in capillaries (10-30 mm long) or in microchips with the same channel length. A 10-30x reduction of the conductivity of the separation medium was observed during the establishment of pH gradient. Results obtained using different IEF voltages indicate that there is a nonlinear relationship between the conductivity of an established pH gradient and the applied electric field. Our theoretical analysis using a simplified model generated values that reasonably agree with the experimental data. In addition, we found that above a certain electric field ( approximately 300 V/cm), resolution does not increase with the applied voltage as predicated; we observed band-broadening and gel breakdown. The approach presented in this work can be used for optimization of the IEF separation and judicious selection of IEF conditions. |
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