SignalkaskadenAssays

SignalkaskadenAssays are a class of laboratory assays designed to measure the activity of cellular signaling cascades. They aim to quantify how extracellular stimuli propagate through networks of receptors, kinases, second messengers, and transcription factors to elicit functional responses within cells.

Principle and readouts: The assays rely on readouts that reflect pathway activity. Common approaches include reporter gene assays in which a pathway-responsive promoter drives a luminescent or fluorescent reporter; phosphorylation assays using phospho-specific antibodies; and biosensor-based assays such as FRET or BRET that report on protein interactions or conformational changes in real time. Some assays measure endpoint outputs such as transcriptional activity, while others track dynamic signaling by live-cell imaging or kinetic luminescence.

Workflow: Experimenters select a pathway of interest, choose an appropriate assay format, and expose cells or lysates to stimuli and controls. After defined incubation, the readout is measured with a plate reader, microscope, or flow cytometer. Data are analyzed to quantify pathway activation, dose–response relationships, and temporal dynamics.

Applications: SignalkaskadenAssays are widely used in basic research to map signaling networks, in drug discovery to identify modulators of specific pathways, in toxicology to assess pathway perturbation, and in systems biology to integrate signaling with downstream responses.

Advantages and limitations: They provide functional, pathway-specific readouts that can be high-throughput. Limitations include context dependence, potential cross-talk between pathways, and the challenge of isolating single-pathway effects in complex networks. Validation with orthogonal methods is common.