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001-es BibID:	BIBFORM022767
Első szerző:	Mocanu, Maria-Magdalena
Cím:	Comparative analysis of fluorescence resonance energy transfer (FRET) and proximity ligation assay (PLA) / Mocanu M. M., Váradi T., Szöllosi J., Nagy P.
Dátum:	2011
ISSN:	1615-9853
Megjegyzések:	Both fluorescence resonance energy transfer (FRET) and proximity ligation assay (PLA) are techniques used in the investigation of protein interactions but the latter has not been evaluated in a systematic way, prompting us to compare their performance quantitatively. Proteins were labeled with oligonucleotide- or fluorophore-conjugated antibodies and their proximity was analyzed by flow cytometry in order to obtain statistically robust data. Both intermolecular and intramolecular PLA signals reached saturation at high expression levels. At the same time, the FRET efficiency was independent of, while the FRET signal exhibited a strict linear correlation with the expression levels of proteins. When the density of oligonucleotide- and fluorophore-conjugated antibodies was systematically changed by competition with unlabeled antibodies the FRET signal was linearly proportional to the amount of bound fluorophore-tagged antibodies, whereas the PLA signal was again saturated. The saturation phenomenon in PLA could not be eliminated by decreasing the duration of the rolling circle amplification reaction. Our data imply that PLA is a semiquantitative measure of protein colocalizations due to non-linear effects in the reaction and that caution should be exercised when interpreting PLA data in a quantitative way.
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban Cell biology Fluorescence resonance energy transfer Flow cytometry Protein associations Proximity ligation assay
Megjelenés:	Proteomics. - 11 : 10 (2011), p. 2063-2070. -
További szerzők:	Váradi Tímea (1982-) (okleveles vegyész) Szöllősi János (1953-) (biofizikus) Nagy Péter (1971-) (biofizikus)
Internet cím:	Szerző által megadott URL DOI Intézményi repozitóriumban (DEA) tárolt változat
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