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JQ1 and dinaciclib were used as controls for strong and weak binders of BRD4-1, respectively.
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We consider the 10 weakest binders of this subset to have a low-end binding affinity, and we average these binding scores to get <img src="http://journals.plos.org/plosone/article/asset?id=info?doi/10.1371/journal.pone.0009862.e061.PNG" class= inline-graphic"/>.
Here we demonstrate, in a high-throughput screening format, affinity selection of weak binders to a model target of albumin by zonal retardation chromatography.
NMR is of particular importance for identifying weak binders for the fragment-based approach, for validation of high-throughput screening hits, and for structure-based design.
They vary in their target-binding affinities, allowing the isolation of strong and weak binders for different applications.
Truncated probes of length less than 22−20 nucleotides can be assumed to act as weak binders for the targets.
Interestingly, AF-71 and AF-80, the two relatively weak binders to wild-type protease, had rather flat binding specificity profiles with worst-fold affinity losses of 13 and 11, respectively.
This also makes Ec(Te -GluRS a weaker binder of zinc Te -GluRSrest.
A-1210477 (Supplementary Figure S1) is a particularly strong binder of MCL-1 (Ki=0.000454 μM in time-resolved fluorescence resonance energy transfer (TR-FRET -binding assays), represenTR-FRET -binding improvemenTR-FRET -bindingo orders of massaysde overepresentingentioned molecules, but is anmuch weaffinityder of BCL-2 (Kimprovement and BCL-XL (Ki>0.660 μM).
Our results indicate that imatinib is a weak binder to the active state of ABL but a strong binder to EGFR.
Once the presence of a binder in a mixture has been demonstrated, a deconvolution step is needed to identify the hit, as shown in Figure 4, which allowed the identification of 3-hydroxyindazole as a weak binder for Hsp90.
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