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In this study, we employed a two-step aptamer selection strategy, comprised of a recombinant protein-based selection from the library followed by a cell-based binding screening for the selection of a thioaptamer to E-selectin.
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Through a cell surface binding screen for FLRT3 partners, we identified Unc5B and Unc5D as high-affinity FLRT3 interactors.
While these probes could be used in competitive binding screens for substrate phosphorylation site inhibitors of PKA or Pim-1, this application has not been reported.
In the multi-receptor binding screen, pIC50 values for ZD4054 at ETA were 22, 27 and 13 n M (mean value 21 n M) (Table 1).
Next, all of the DNA sequences near sites of Tup1 binding were screened for the presence of DNA sequence motifs that corresponded to the specificities of each of the recruiters [7], [31] (Figure 2B).
In the previous work, a variety of fluorophores were attached at two different sites on E. coli SSB and the fluorescence increase upon ssDNA binding was screened for each combination (Dillingham et al., 2008).
After three rounds of panning, 24 clones of the second round and 23 clones of the third round of panning were picked randomly for antigen-binding screening.
Screening for binding site similarity to the ATP-binding site of bovine synapsin (PDB entry 1aux) was done as described above.
Extensive screening for binding to other neuroreceptor subtypes demonstrated their P2X7 selectivity.
For example, screening for binding elements in RNA sequences comparable to the approach implemented for the PTBs, Nova, and Pumilio in this study could be readily performed for any RBP of interest.
At the heart of the screening for fragments binding to a specific target, crystallography delivers structural information essential for subsequent drug design.
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