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PathHunter® eXpress TORC1 Nuclear Translocation Assay

PathHunter® Nuclear Translocation cells are engineered to co-express two fusion proteins: a) Enzyme Donor (ED) tagged target protein; b) an Enzyme Acceptor (EA) tagged TAZ domain, derived from the CBP/P300 transcription factor, that localizes to the nucleus. Depending on the assay, activation of the signaling pathway can either a) induce translocation of the ED-tagged target protein into the nucleus, which will force complementation of the two enzyme fragments, and result in the formation of a functional enzyme that will hydrolyze substrate and generate a chemiluminescent signal; or b) induce the ED-tagged protein to vacate the nucleus, resulting in a decrease of functional enzyme and a subsequent decrease of chemiluminescent signal. Some nuclear translocation assays will also co-express an untagged secondary protein involved in the pathway of interest. These cells have been modified to prevent long term propagation and expansion using a proprietary compound that has no apparent effect on assay performance.

Catalog No. Size Price Quantity
93-0833E3CP19S 100 dp (1 x 96-well) $1378.00
93-0833E3CP19M 200 dp (2 x 96-well) $2058.00
93-0833E3CP19L 1000 dp (10 x 96-well) $7155.00

For account specific pricing, please contact Customer Service at customerservicedrx@eurofins.com.
Target/Description
CREB regulated transcription coactivator 1
Group
Transcription factor
Cell Background
U2OS
Species
Human
Accession Number
NM_015321.1
Agonist/Ligand
Forskolin
Assay Measures
TORC1 Translocation
Cells were plated in a 96-well plate and incubated at 37°C/5% CO2. Cells were then stimulated with a control compound, using the assay conditions described below. Following stimulation, signal was detected using the PathHunter Detection Reagents provided in the kit according to the recommended protocol. For a detailed protocol, please refer to the user manual.
Format
See User Manual for Product Configuration Details
Storage
Short term (<24 h): Store at -80°C; Long term (>24 h): Store in vapor phase of liquid nitrogen.
Coupling
N/A
  • [User Manual] PathHunter® eXpress Nuclear Translocation Assays

    PathHunter® eXpress Nuclear Translocation assay kits provide a robust, highly sensitive and easy-to-use, cell-based functional assay for monitoring target protein translocation into the nucleus upon activation of the pathway. The eXpress kits contain everything needed for a complete experiment including cells, detection reagents, media and plat...

    [User Manual] PathHunter® eXpress Nuclear Translocation Assays