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Not to mention, when a song is mastered the tones and overtones are 'equalized' to control unwanted subharmonics and noises, such as fret buzz and finger-drag.
The relatively low acceptor quantum yield will limit to some extent the use of the FRET couples described in this study for application in acceptor-based FRET applications such as FRET based biosensors, of which cameleon [42], [43] is the best-known example.
Cyan fluorescent protein (CFP) was used as FRET donor and yellow fluorescent protein (YFP) as FRET acceptor.
We plotted FRET measurements from metaphase and anaphase kinetochore clusters separately, as FRET was found to be systematically higher in anaphase.
FRET efficiencies were calculated from integrated donor (I D) and acceptor (I A) intensities as FRET = I A/(I D + I A 33,34.
The FRET signals were directly visualized under the microscope and quantified as FRET concentration (FRETc) with the Olympus FRET analysis program19,20.
Lectures and primary literature critiques on topics such as fluorescence microscopy, including applications such as FRET and single-cell analysis; human and murine genetic analysis; FACS; proteomics and analysis of noncoding RNAs.
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Substrate cleavage was readily monitored and was linear for up to 4 h but enzyme concentrations of about ten-fold higher were required compared to assays using protein substrate LC3 or analogs thereof (such as FRET-LC3).
As FRET-positive control we generated the pEYFP-ECFP construct expressing EYFP and ECFP as a fusion.
We employ a molecular imaging methodology, such as FRET-based microscopy, to show that this probe allows monitoring, and accurately reflects consistent structural changes of the PS1 molecule in live cells, and that these changes closely correlate with changes in the Aβ42/40 ratio.
Evidence for direct interaction of MamJ and MamK comes from two-hybrid assays as well as FRET-experiments.
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