May 2009
This application note describes how the ActiPix UV area imaging detector can be used to test biocatalyst substrate specificity towards a mixture of UV active compounds using a continuous engagement electrophoretically mediated microanalysis (EMMA) assay method.
A plug of the putative substrates is injected into a fused silica capillary containing the background electrolyte and dissolved enzyme. The components are separated in the initial part of the run, avoiding the competitive binding and inhibition problems that can occur in standard enzyme assays. The peaks corresponding to each of the separated components are measured at multiple time points along the looped capillary, providing intrinsic self-referencing and allowing the identification of good substrates as those components that show decreased peak areas over time.
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