Optimizing Parameters for Hands Free cIEF

cIEF analysis requires the protein sample to be pre-mixed with carrier ampholytes, pI markers, and other additives. Proteins can be sensitive to highly basic environments and can experience degradation when exposed to these conditions for extended periods. The iCE3 automates sample preparation minimizing protein sample degradation in IEF buffers and operator to operator variability. The sample is prepared immediately prior to injection limiting sample exposure to cIEF buffers and preventing degradation. The development of a robust and repeatable automated sample preparation assay requires optimizing sample mixing parameters. During optimization, several factors need to be considered, such as the shape of the sample vial, the number of mixing strokes, air bubbles and sample dilution by the mixing action.