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PathHunter® Toolbox Assays
The PathHunter® tool box approach allows you to build your own assay based on Enzyme Fragment Complementation (EFC) for protein translocation. It has been successfully applied to a variety of targets and signaling events including nuclear hormone assays, pathway indicators as well as endocytosed receptors. Tool-Box or Open system are stable clonally-derived cell lines, which can be utilized to introduce your protein of interest with a ProLabel™ or ProLink™ EFC tag. This protein can then be monitored as it translocates into a nucleus or endosome or cytosol. Simple, mix-and read protocol with a variety of complimentary detection reagents, antibodies now offers a complete solution for your proprietary or intractable targets.
Product Features
Versatile
Build your own EFC assay
Append your protein of interest to ProLink or ProLabel
Compatible with a variety of detection reagents and standard
benchtop luminometers
Results in simple one-step chemiluminescent assay
How do I build my own assay?
You would need
1. PathHunter Parental cells
2. Vectors
3. Detection reagents
4. ProLink/ProLabel Antibody
5. EAstern Kit
Click here to view an assay flowchart.
Click on figure to enlarge
Figure 1. PathHunter® EA-Nuc parental cells were used to build P53 translocation cell line. P53 was tagged to ProLabel and the data shows P53 trasnslocation can be measured in response to compounds that affect DNA damage pathway in a simple chemiluminescent gain of signal assay.
Click on figure to enlarge
Figure 2. Expression of EA restricted to the cytoplasm. Confocal images showing immuno-labeling of the EA protein using PathHunter Anti-EA Antibody (PN, 93-0246) in the PathHunter CHO-K1 EA-Cyto Cell Line demonstrating the specific localization of the EA protein within the cytoplasm and the homogeneous expression of EA in the clonally derived cell population.
PathHunter® Parental Cell Lines
Click here to view reagent calculator.
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