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Fractions showing the highest fluorescence change upon ADP binding were pooled, and the labeled protein was concentrated in an Amicon Ultracentrifugal filter device (Millipore Corporation) to ∼10 mg mL−1.
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[Ca2+] signals were detectable at a high signal-to-noise ratio because of the high relative fluorescence change of G-CaMP2 upon [Ca2+]-binding [4].
Called SPARK (separation of phases-based activity reporter of kinase), these reporters have large dynamic range (fluorescence change), high brightness, fast kinetics, and are reversible.
It can display on off type fluorescence change with high selectivity and sensitivity toward Fe3+ in DMF H2O solution.
Studying the fluorescence change with higher time scale resolution would reveal the depolarization events secondary to hemichannel opening.
The decrease in the magnitude of the fluorescence change at higher concentrations of the amino acid suggests that binding at the active site may be associated with an additional change in the environment of Trp120.
We used the highest fluorescence plateaus reached to determine the maximal fluorescence change, which in some cases were from the 62 Hz trains since not all cells could always follow the 82 Hz stimulation reliably.
The latter suggestion is more plausible because it was found that the amplitude of the fluorescence change decreased at higher concentrations of dATP (data not shown).
A remarkable fluorescence change is observed with high selectivity toward OTBs over organotin fluorides, organotin chlorides, ionic bromides, or other covalent organobromines.
GCaMP3 has increased fold fluorescence change (increased Fmax/Fmin) and higher Ca2+ affinity (660±19 nm) than many other genetically encoded indicators.
Measurements of DNA binding to DCC-SSB suggest that at low salt, the fluorescence change is ∼75% that at high salt (19).
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