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The identity of the protein and the specificity of the binding were verified by competition with a point mutated mb-1/EBF site and by the inclusion of an EBF specific antibody into the reaction mixture.
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Islet binding was verified by immunostaining the same sections to detect insulin.
In parallel studies, the specific cell uptake due to σ receptor binding was verified by the presence of a variety of σ ligands including -pentazocine (σ1), haloperidol (non-selective σ1/σ2) and unlabelled SIG343 and SIG353 (σ2).
For each of the peptides HLA-A*0201 HLA-A*0201s verified bindingmpetition assay.
The presence of anti-CD15-SPIONs binding was verified by Prussian blue staining.
Concentration-dependent binding was verified by a linear response when plotting on double-logarithmic scale.
The specificity of aptamer-STIV binding was verified by electrophoretic mobility shift assay (EMSA), as described previously [ 20– 20].
The effect on miRNA binding was verified by computing the difference in hybridization energy (ΔΔG) caused by that particular SNP (see Materials and methods section).
Not only the binding sites were verified by an extensive mutational analysis [10], but now the conformational changes were also observed in solution by NMR spectroscopy.
Computational predictions of FOXO and E-box binding sites were verified by ChIP assays.
The binding sites were verified by SYBR® Green Based qPCR using a CFX connect™ real-time PCR detection system (Bio-Rad, Richmond, CA).
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