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The fluorescence of -1EF+2EF enhances ~20% upon Ca2+ binding and 50 μM Ca2+ is enough to saturate the binding (data not shown).
The binding of E4.3 and J4.48 clone was comparably diminished (J4.48 binding data not shown).
Deletion of activators did not effect the apparent level of Taf1 binding (data not shown).
Furthermore, inhibiting neddylation by expressing a dominant negative form of Ubc12 does not increase CAND1 binding (data not shown).
Although repeats 7 and 8 are not necessary for binding, they are not inactive in binding (data not shown).
Furthermore, the ZF domains of BORIS isoproteins are both sufficient and necessary for DNA binding (data not shown).
Cellular viability was confirmed using a CyQuant NF assay, which measures cellular content via fluorescent dye binding (data not shown).
The mutation of individual binding sites showed no effect on PTB binding (data not shown, Fig 4a and b).
Munc18-1 withethe Glu379 mutation did not show any evidence for increased Rab3 binding (data not shown).
Gel shift experiments with recombinant dTCF demonstrated that the introduced mutations eliminated dTCF binding (data not shown).
L. interrogans LigB, which binds both the NTD and the GBD [3], served as a positive control for GBD binding (data not shown).
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Since I tried Ludwig back in 2017, I have been constantly using it in both editing and translation. Ever since, I suggest it to my translators at ProSciEditing.

Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com