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A TF may bind to a site on the DNA but this binding event is not necessarily functional.
Moreover, label-free methods for glycoprofiling are of particular interest, because the binding event is not compromised by the label.
This high-affinity binding event is not discerned during AUC because the mass change induced by binding the 20 kDa protein to the 169 kDa duplex is small.
With some notable exceptions [ 37], simulating a receptor-ligand system long enough to capture an entire binding event is not currently feasible.
The number of tags expected to map to a binding event is not only dependent on the binding level at a particular site, but also the composition of the total binding event population that is being sampled.
Overall, our results suggest that the combinatorial action of a group of transcription factors is responsible for the induction of a small subset of CREB-bound promoters in the liver, and the simple presence of a CRE or even a proven CREB binding event is not sufficient to determine whether the associated gene is part of the fasting response.
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At high salt conditions the second binding event was not apparent (Fig. 4D) and fitting the data to a tight binding quadratic equation, produced relatively good fits with R values of 0.9354 and 0.9445 and Kd values of 9.4 and 36 nM for dT70 and dT35, respectively.
Alternatively, the two binding events are not necessarily sequential.
Additionally, there is no consistent arrangement of the Adr1 and Cat8 binding sites within promoters of co-regulated genes [9], supporting the conclusion that two separate activator binding events are not necessary for activation of these genes.
Many transcription factor binding events are not associated with gene activation when assessed on a genome-wide scale.
As noted above, these subsequent binding events are not detected in the EMS studies except at IHF concentrations of >1 μM.
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