Translocation Assay: Translocation of NFΚB under three different conditions. In condition A (left picture) NFΚB is mainly cytoplasmic while condition B (middle picture) has led to a shift of NFΚB to the nucleus. These images are easily quantified by High Content Analysis and an example time course shown (right panel). Condition A is shown in yellow while B is in red. Note a further condition also induced NFΚB (shown in green, right picture) but this condition led to a more sustained NFΚB response. Various positive and negative control conditions are shown by the white bars to the right of the time course.

Translocation Assay

Not only can High Content Analysis give you quantification of antibody staining, but it can also give you information about the position of the stain within the cell. This makes the technique ideal for signalling events which result in the movement of a protein from the cytoplasm to the nucleus. Often it is possible to combine a phosphorylation readout with a translocation readout to get even more information about a particular signal transduction pathway. Like flow cytometry, all these measurements can be combined with subpopulation gating to give detailed understanding of cell population heterogeneity.

Courtesy of M Milward and P Cooper, The University of Birmingham, UK