New optical arrangement for CE/“in instrument” LIF analysis, application to protein analysis using v

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  DS PAGE is a widely used analytical method for protein analysis.The visualization of the separated proteins is generally accomplished by staining with either Coomassie Brilliant Blue when extreme sensitivity is not needed or with the more sensitive silver stain dyes.While Coomassie Brilliant Blue is used for semiquantitative evaluation of the relative intensity of the major protein bands, its limit of detection is approximately 100 fold less sensitive than silver staining.Sliver staining methods can improve the limit of detection to 10 ng/mL.One problem using capillary electrophoresis (CE) for protein study is the lack of sensitivity and selectivity with conventional UV-VIS absorption detectors.CE with laser induced fluorescence detection can provide for better selectivity and increase the sensitivity.In this presentation we will show data from a new LIF detector which can be used with all commercial CE instruments.We developed a new optical arrangement using a ball lens for excitation and an ellipsoid for collection of the fluorescence.Optimization of injection using different concentrations of lithium dodecyl sulfate or sodium dodecyl sulfate will be presented.Sensitivity data with LIF versus native fluorescence will be presented.
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