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To analyze binding specificity, we performed free energy calculations of various combinations of RVDs and bases using Poisson Boltzmann surface area (PBSA) and other approaches.
The ability to rapidly select and analyze binding interfaces, and compatibility with high-throughput methods under diverse conditions, makes it likely that the combination of phage display and synthetic combinatorial libraries will prove to be the method of choice for synthetic binding protein engineering for broad applications.
Using this mutant, immunoprecipitation experiments were performed to analyze binding between several β-subunits and mutant bestrophin-1.
STAP is an effective method to analyze binding site arrangements, TF cooperativity, and TF target genes from genome-wide TF-DNA binding data.
To this end we used fluorescence resonance energy transfer (FRET) which we have used previously to analyze binding of HIF-1α to HIF-1β [54] and to prove interaction of PHD2 with the cis/trans prolyl isomerase FKBP38 [30].
Different from existing thermodynamic models, STAP explicitly expresses the expected number of TFs bound to a regulatory sequence, and thus it is directly applicable to analyze binding intensities reflected in whole-genome binding data.
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In this work we analyzed binding of two peptides, namely temporin B and TB_KKG6A, to Escherichia coli cells and to Escherichia coli LPS.
This has triggered the requirement for effective high-throughput screening platforms for analyzing binding by larger chemical libraries to STAT proteins.
In order to answer the question of how binding affinity and specificity is achieved, we analyzed binding energetics on the molecular level, with reference to the available structural data.
We first analyzed binding of relevant trans-acting factors to the β-globin locus.
Similarly, we analyzed binding of a GST-TR fusion to the predicted binding sites of these T3 responsive genes.
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