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, 6 (11), 1748-60

Applying Label-Free Dynamic Mass Redistribution Technology to Frame Signaling of G Protein-Coupled Receptors Noninvasively in Living Cells

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Applying Label-Free Dynamic Mass Redistribution Technology to Frame Signaling of G Protein-Coupled Receptors Noninvasively in Living Cells

Ralf Schröder et al. Nat Protoc.

Abstract

Label-free dynamic mass redistribution (DMR) is a cutting-edge assay technology that enables real-time detection of integrated cellular responses in living cells. It relies on detection of refractive index alterations on biosensor-coated microplates that originate from stimulus-induced changes in the total biomass proximal to the sensor surface. Here we describe a detailed protocol to apply DMR technology to frame functional behavior of G protein-coupled receptors that are traditionally examined with end point assays on the basis of detection of individual second messengers, such as cAMP, Ca(2+) or inositol phosphates. The method can be readily adapted across diverse cellular backgrounds (adherent or suspension), including primary human cells. Real-time recordings can be performed in 384-well microtiter plates and be completed in 2 h, or they can be extended to several hours depending on the biological question to be addressed. The entire procedure, including cell harvesting and DMR detection, takes 1-2 d.

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References

    1. Proc Natl Acad Sci U S A. 2003 Sep 30;100(20):11406-11 - PubMed
    1. Cell Death Differ. 2008 Jan;15(1):134-42 - PubMed
    1. Trends Pharmacol Sci. 2007 Aug;28(8):407-15 - PubMed
    1. J Med Chem. 2008 Nov 27;51(22):7061-4 - PubMed
    1. Mol Pharmacol. 2006 Apr;69(4):1441-53 - PubMed

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