PathHunter™ High-throughput Assays for Cellular Protein Trafficking New! ß-Arrestin Target Data Site
PATHHUNTER™ ASSAYS: WINNER OF THE 2006 FROST & SULLIVAN TECHNOLOGY INNOVATION AWARD!
Overview & Description PathHunter™ products comprise a family of simple, robust cell-based assays and related tools designed to directly measure protein translocation, secretion, degradation, expression, and cell proliferation-without imaging or antibodies. These products may address cell-signaling cascades in which a target protein undergoes nuclear translocation, degradation, or secretion. A variety of targets can be measured, including transcription factors, nuclear hormone receptors, and kinases. Other unique PathHunter assays enable measuring mitotic activity in mammalian cells.

In PathHunter assays, a target protein fused to a ß-galactosidase (ß-gal) peptide (ProLabel™ tag) is expressed in the cytoplasm. Another portion of ß-gal (Enzyme Activator, or EA) is expressed only in the nucleus. When the two interact, active enzyme is formed generating signal. This method can detect protein translocation to the nucleus or nuclear envelope degradation during mitosis.

Key Features
• Homogeneous, chemiluminescent pathway profiling assays
• Precise, robust assays with high S:B
• No imaging, special equipment, antibodies, or GFP required
• No license fee

Homogeneous Antibody-free Assays
PathHunter products feature a novel, non-imaging approach based upon detecting complementation of two ß-galactosidase fragments, which generates a functional enzyme and a chemiluminescent signal. Consequently, no antibody labeling or imaging is required, and the assays are compatible with standard microplate readers. The basic PathHunter procedure is elegantly simple, and its homogeneous, two-step format offers high-sensitivity chemiluminescent detection.

PathHunter™ Assay Protocol

Comparison of Pathway Profiling Technologies

Ready-to-use Assays or Custom Targets
For maximal convenience, PathHunter technology include variety of options that allow one to develop and study assays for target proteins known to undergo translocation, secretion and degradation.

PathHunter Biosensor Cell Lines are double-stable cell lines with the EA ß-gal fragment plus one of a variety of targets expressed as a ProLabel-tagged fusion protein.

PathHunter Custom Assay Development allows one to assay virtually any target known to undergo translocation.