%0 Journal Article %1 Manoharan2015 %A babu Manoharan, G. %A Enkvist, E. %A Kasari, M. %A Viht, K. %A Zenn, M. %A Prinz, A. %A Filhol, O. %A Herberg, F. W. %A Uri, A. %D 2015 %J Analytical Biochemistry %K herberg myown %P 10-17 %R http://dx.doi.org/10.1016/j.ab.2015.04.009 %T FRET-based screening assay using small-molecule photoluminescent probes in lysate of cells overexpressing RFP-fused protein kinases %U http://www.sciencedirect.com/science/article/pii/S0003269715001554 %V 481 %X Abstract An assay was developed for characterization of protein kinase inhibitors in lysates of mammalian cells based on measurement of \FRET\ between overexpressed red fluorescent protein (TagRFP)-fused protein kinases (PKs) and luminophore-labelled small-molecule inhibitors (ARC-Photo probes). Two types of the assay, one using TagRFP as the photoluminescence donor together with ARC-Photo probes comprising red fluorophore dyes as acceptor, and the other using TagRFP as the acceptor fluorophore in combination with a terbium cryptate-based long-lifetime photo-luminescence donor, were used for FRET-based measurements in lysates of the cells overexpressing TagRFP-fused PKs. The second variant of the assay enabled the performance of the measurements in time-resolved conditions that lead to substantially higher values of the signal/background ratio and further improved the reliability of the assay.

@article{Manoharan2015, abstract = {Abstract An assay was developed for characterization of protein kinase inhibitors in lysates of mammalian cells based on measurement of \{FRET\} between overexpressed red fluorescent protein (TagRFP)-fused protein kinases (PKs) and luminophore-labelled small-molecule inhibitors (ARC-Photo probes). Two types of the assay, one using TagRFP as the photoluminescence donor together with ARC-Photo probes comprising red fluorophore dyes as acceptor, and the other using TagRFP as the acceptor fluorophore in combination with a terbium cryptate-based long-lifetime photo-luminescence donor, were used for FRET-based measurements in lysates of the cells overexpressing TagRFP-fused PKs. The second variant of the assay enabled the performance of the measurements in time-resolved conditions that lead to substantially higher values of the signal/background ratio and further improved the reliability of the assay. }, added-at = {2015-04-14T15:46:03.000+0200}, author = {babu Manoharan, G. and Enkvist, E. and Kasari, M. and Viht, K. and Zenn, M. and Prinz, A. and Filhol, O. and Herberg, F. W. and Uri, A.}, biburl = {https://www.bibsonomy.org/bibtex/2d33f78ddf29c90103f3eea3bb7c20dc0/biochemie}, description = {FRET-based screening assay using small-molecule photoluminescent probes in lysate of cells overexpressing RFP-fused protein kinases}, doi = {http://dx.doi.org/10.1016/j.ab.2015.04.009}, interhash = {d04cbc0678fac577ad5a169960bcf58f}, intrahash = {d33f78ddf29c90103f3eea3bb7c20dc0}, issn = {0003-2697}, journal = {Analytical Biochemistry}, keywords = {herberg myown}, pages = {10-17}, timestamp = {2015-07-27T16:01:23.000+0200}, title = {FRET-based screening assay using small-molecule photoluminescent probes in lysate of cells overexpressing RFP-fused protein kinases}, url = {http://www.sciencedirect.com/science/article/pii/S0003269715001554}, volume = 481, year = 2015 }