PathHunter™ Cell Cycle Assays
Overview & Description
PathHunter™ Mitotic Index Assays are novel biosensor cell assays that uniquely meet the need for easy, cost-effective HTS assays to identify compounds or biologicals like siRNAs that inhibit mammalian cell proliferation. PathHunter™ Cell Lines and Detection Kits are used together to detect nuclear envelope degradation at mitosis.
The ready-to-use assays advance screening for new cancer therapeutics because, unlike other mitotic assays that require complex and expensive HCA imaging, flow cytometry or antibodies, PathHunter Assays are homogeneous, without antibodies, and results are obtained using standard plate readers. HEK 293 (human embryonic kidney) and CHO-K1 (hamster) double-stable cell lines are currently available.
Biosensor Cells: A Breakthrough Enabling HTS
These HEK 293 and CHO-K1 biosensor cells uniquely combine a homogenous format with the in vivo mammalian cellular context needed to rapidly identify quality anti-cancer hits. Furthermore, mitotic inhibition in cultured cells increases chemiluminescence, providing positive signal gain, rather than the negative read-out common in other assays for anti-proliferatives.
Anti-cancer Agent Discovery & Characterization
PathHunter Mitotic Index Assays enable assessment of mitotic activity in cells treated with anti-cancer test compounds or siRNAs. The assays are sensitive enough to differentiate between inhibitors of mitosis and other cell cycle phases, such as DNA replication in S phase. Validation studies also show they positively identify mitotic inhibitors among known anti-cancer compounds, with low false positives.
Features and Benefits
» Advanced, Homogenous Assay Format
• Enables whole-cell, functional assays
• Quickly identifies mitotic inhibitors by signal gain
• Also detects non-mitotic inhibitors with signal loss
» Easy to Use
• Just two steps-add test compound and detection reagent
• Needs no fixation, imaging, or antibody labeling
• Compatible with automated screening
» Chemiluminescent Signal
• Uses any standard plate reader
• Minimizes compound interference
ß-galactosidase Complementation Detects Cell Division
PathHunter™ Cell Lines are engineered with nuclear and cytoplasmic fragments of ß-galactosidase. At mitosis the nuclear envelope degrades and allows the ß-gal fragments to complement, forming active enzyme that cleaves a chemiluminescent substrate provided in the PathHunter EFC Detection Kit (purchased separately).
» Double-stable Mitotic Index PathHunter Cell Line
» PathHunter Detection Kit purchased separately
» Luminometer, Multimode Microplate Reader or CCD Camera