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For the dissociation rate constant analysis, complete initial binding is confirmed by a gel shift, and then a 100-fold excess of unlabeled DNA 24-mer is added.
The importance of electrostatics in MBD binding is confirmed by the ability of the poly dA-dT) compoly dA-dT ameliorate the influencompetitor.
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Plaque binding was confirmed by thioflavin-S staining.
Cell binding was confirmed by visualizing adherent cells by fluorescence microscopy.
The EstC-NH2 grafting on the thiol MUA spacer through the formation of an amide binding was confirmed by the PM-RAIRS and XPS complementary experiments.
The specificity of the binding was confirmed by inhibition studies, which showed reduced crosslinking signal after pre-incubation of C3 with compstatin but not with various inactive analogs.
The essential role of Phe313 and Trp305 in N-6 binding was confirmed by our SPR assays, showing that N-6 had much lower binding affinity to two RXRα-LBD point mutants with Ala substitution of Phe313 or Trp305 (Fig. 4D and 4E).
The specificity of [18F]FEDL binding was confirmed by pre-blocking of the tissue sections with excess of non-radiolabeled lactose.
The in vitro binding was confirmed by overlay of the blotted membranes with radio-labeled HAMLET (Figure 2A, right panel).
The specificity of this binding was confirmed by competition studies using cold wild type and mutant competitor probes.
All peptides were less than fifty amino acids in length and the binding was confirmed by in vivo as well as in vitro studies.
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