Expression systems readout

Expression systems with direct or indirect fluorescence readout... 

While dispersion is defined as the physical dilution process of the injected sample zone in the FIA system, the dispersion coefficient D is a numerical expression of the dispersion process in a particular element of fluid of the dispersed sample zone, being defined as the ratio of the concentrations before and after the dispersion process has taken place in that element of fluid that yields the analytical readout, that is, D = C°/C, which for C = yields D = where Co is the original concentration of the injected sample... 

Fig. 2 Increasing content recovery by coupling luciferase-based assays with high-throughput biochemical readouts. The same cell line subjected to the luciferase-based assay protocol (HCT116 cells) is evaluated here for its reliability In reporting a biochemical readout (p53 expression) by dot blot analysis. Cells transfected with indicated expression construct and pathway reporters in a 96-well culture plates were lysed 48 h posttransfection. Following luciferase activity measurements (data not shown), protein from spent lysates was immobilized on nitrocellulose using a liquid handler and filtration manifold, (a) p53 and p-actin protein levels detected using protein-specific antibodies, infrared fluorescent dye-coupled secondary antibodies (with emissions at 680 and 800 nM), and the Li-COR imaging system. Columns of lysate corresponding to cells transfected with p53 DNA are boxed, (b) Quantification of the p53 to p-actin protein ratio...

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