Novel therapeutics need to affect their targets in a cellular background. However, cell based assays are often more expensive and difficult to automate. The DiscoverX PathHunter® provides a technology that overcomes these limitations if miniaturized. It reports on stimulation of cellular G-protein coupled receptors and cellular receptor kinases.
PathHunter reporter cell lines and stimulation reagents were added to 1536 or 3456 well plates using the Echo® 555 liquid handler. Labcyte's liquid handler uses an acoustic method for dispensing low volumes. The chemiluminescent signal of the reporter cell lines was acquired by a PHERAstar® microplate reader.
The stimulation of either tropomyosin receptor kinase A by β-NGF, of PRLR-JAK pathway by prolactin or of the dopamine receptor 2 with pergolide resulted in an increase in signal derived from the dedicated reporter cell line. As few as 1,000 cells in a 1536 well plate or 250 cells in a 3456 plate were sufficient to detect pathway stimulation.