Applying label-free dynamic mass redistribution technology to frame signaling of G protein–coupled receptors noninvasively in living cells

Schröder, R. et al. (2011) Applying label-free dynamic mass redistribution technology to frame signaling of G protein–coupled receptors noninvasively in living cells. Nature Protocols, 6(11), pp. 1748-1760. (doi: 10.1038/nprot.2011.386)

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Publisher's URL: http://dx.doi.org/10.1038/nprot.2011.386

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<sup>2+</sup> 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

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Milligan, Professor Graeme
Authors: Schröder, R., Schmidt, J., Blättermann, S., Peters, L., Janssen, N., Grundmann, M., Seemann, W., Kaufel, D., Merten, N., Drewke, C., Gomeza, J., Milligan, G., Mohr, K., and Kostenis, E.
College/School:College of Medical Veterinary and Life Sciences > Institute of Molecular Cell and Systems Biology
Journal Name:Nature Protocols
ISSN:1754-2189
ISSN (Online):1750-2799

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