The PathHunter
® GPCR Explorer Kit is a simple and easy-to-use method for building on-demand functional GPCR assays in just 3 simple steps – Clone. Transfect. Read. Whether you have a panel of mutant GPCR cDNAs or a favorite GPCR heterodimer you’d like to functionally test, the simple, optimized transfection protocol combined with PathHunter
® EFC-based chemiluminescent detection makes it the ideal platform for exploring novel GPCR receptor biology.
Features & Advantages
- Based on G-protein independent β-Arrestin read-out
- Contains highly transfectable, ready-to-assay CHO-K1 cells
- Choice of vectors optimized for maximal expression in mammalian cells
- Ideal for GPCR heterodimerization, siRNA knockdown, or mutant receptor studies
- Chemiluminescent detection used with any standard luminometer
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How it Works |
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Assay Performance
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Create Customized GPCR Biosensor Lines
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CHO-K1 GPCR eXplorer cells were plated at 10,000 cells per well and incubated overnight at 37°C. The following day, cells were transfected with 2 µg of pEGFP vector and 48 hr post-transfection, greater than 60% of the GPCR Explorer cells were transfected ([Left panel) [click to enlarge]. In a separate experiment, eXplorer cells were transiently transfected with 2 µg of the pCMV-SSTR2-PK vector (+ SSTR2 DNA) or an empty vector (- SSTR2 DNA). Twenty-four hours later, cells were treated with increasing concentrations of Somatostatin 28, a control agonist. Activation of the SSTR2 receptor stimulates binding of Arrestin resulting in an increase in β-gal enzyme activity that can be measured using chemiluminescent PathHunter Detection Reagents (Right panel) [click to enlarge].
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