PathHunter™ Nuclear Translocation Assays are single-addition, homogeneous assays that measure the effects of agonist or antagonist compounds on target protein translocation in vivo to the nucleus. The assays facilitate cost-effective, high throughput screening, because they are simple, use microtitre plates, and do not require imaging or antibodies. A PathHunter CHO-K1 Cell Line expressing a protein known to translocate to the nucleus, such as the glucocorticoid receptor, is used together with the PathHunter Detection Reagent Kit for chemiluminescent read out.
Features and Benefits
» In vivo Whole Cell Assay
• Provides a high-throughput protein translocation assay in a simple, cost-effective format
» No Imaging, Antibodies, or GFP
• Reduces assay development, data analysis and data storage
» No Fluorescent Interference
• Eliminates optical interference from fluorescent compounds by chemiluminescent format
• Provides mix and read protocols to facilitate assay development and automation
• Uses standard microplates, existing liquid handlers, and a wide range of luminometers, mutlimode microplate readers or CCD devices.
Measuring Protein Translocation in Whole Cells using PathHunter EFC Detection
Ready-to-use Biosensor Cell Lines stably express the Enyme Acceptor (EA) ß-galactosidase (ß-gal) fragment exclusively in the cell nucleus and the target of interest in the cytoplasm as a ProLabel fusion protein. Upon translocation of the cytoplasmic protein to the nucleus, ß-gal activity is restored and a chemiluminescent signal is generated.
» Double-stable PathHunter Cell Line for a specific target
» PathHunter EFC Detection Reagents purchased separately
» Luminometer, Multimode Microplate Reader or CCD Camera