Robust Assays in Living Cells
» Easy assay steps – No cell lysis
» BacMam delivery to most cell types
» Fluorescence imaging or plate reader detection (Z’ > 0.8)
» Physiologically-relevant and cell friendly
» No forskolin or IBMX needed
» No enzymes or co-factors
» Multiplex GPCR Assays: Gs, Gq, Gi, Voltage
Simultaneous measurements of GPCR signaling in live HEK 293 cells. Green sensors in the top panels, red sensors in the bottom panels. M1 muscarinic receptor activated by carbachol.
Recent discoveries using our live cell fluorescent sensors:
- Q. Wang, A. N. Pronin, K. Levay, J. Almaça, A. Fornoni, A. Caicedo, V. Z. Slepak. Regulator of G protein signaling Gβ5-R7 is a crucial activator of muscarinic M3 receptor-stimulated insulin secretion. FASEB J. 2017 Jul 7, fj.201700197RR.
- P. Tewson, S.Martinka, C. Berlot, T. Hughes, A.M. Quinn, S. Tillo, Measuring dynamic and real-time cAMP levels using cADDis, a live-cell indicator for Gs and Gi signaling. SLAS 2017.
- SLAS Conference Abstract, 2017. Direct Measurements of Gi signaling and its dynamic interplay with Gs using cADDis, a live-cell assay for cAMP with improved sensitivity.
- P. Tewson, A.M. Quinn, T. Hughes. Dynamic Measurements of Gi Signaling in Living Cells. FASEB J. 2016 April, vol. 30 no. 1. Supplement lb478
- J. Almaça, J. Molina, D. Menegaz, A.N. Pronin, A. Tamayo, V. Slepak, P. Berggren, A. Caicedo. Human beta cells produce and release serotonin to inhibit glucagon secretion from alpha cells. Cell Reports, Volume 17, Issue 12, p3281–3291, 2016.
- GLP-1 stimulates insulin secretion by PKC-dependent TRPM4 and TRPM5 activation. J Clin Invest. 2015; 125(12):4714-4728.
- Does PKC activation increase the homologous desensitization of µ opioid receptors? Br J Pharmacy. 2015; 172(2):583-92.
- PKC-dependent Phosphorylation of the H1 Histamine Receptor Modulates TRPC6 Activity. Cells. 2014. 3(2): 247-257.
- New DAG and cAMP Sensors Optimized for Live-Cell Assays in Automated Laboratories. J Biomol Screen. 2015. DOI: 10.1177/1087057115618608
- A multiplexed fluorescent assay for independent second-messenger systems: decoding GPCR activation in living cells. J Biomol Screen. 2013. 18, 797–806Simultaneous Detection of Ca2+ and Diacylglycerol Signaling in Living Cells. PLoS ONE. 2012. DOI: 10.1371/journal.pone.0042791
- Cilia Have High cAMP Levels That Are Inhibited by Sonic Hedgehog Regulated Calcium Dynamics. PNAS 2016. In Press
Fluorescent Assays for Live Cell Discovery