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The cells were then re-suspended in 50 µl 0.5%BSA/PBS containing a 1 50 dilution of sheep anti-human γ chain (FITC) fluorescein isothiocyanate-conjugated antibody (The Binding Site) was added to each well.
A ribosome binding site was added by PCR and is shown in boldface type.
To determine if the SH2 binding is tyrosine site dependent, a synthesized phosphopeptide corresponding to EGFR tyrosine 1068, containing the Grb2 SH2 consensus binding site, was added as a blocker.
If excess competitor DNA containing a seven-nucleotide mutation of the BP1 binding site was added (mbcl-2), however, little competition for binding was observed (lanes 5 and 6).
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Six tandem copies of a synthetic miR-312 binding site were added to the 3' UTR of a UAS-GFP transgene (Fig. 1C).
A bis-ANS solution (10 μmol, equal to twice the amount required to saturate the binding sites) was added to a 0.1 mg/mL peptide solution in phosphate buffer (pH 7.2).
To confirm the specificity, competition using 30-fold excess unlabelled oligonucleotides containing consensus or mutated STAT3 binding sites were added for 20 min at room temperature, after incubation with the radio-labelled oligonucleotides.
Also as expected, we observed a higher level of enrichment with the REP protein as opposed to the FL mutant when the putative TBPH binding site (UG 9 was added to head extract samples (Figure 6A).
Since Mg2+ ions are known to be involved in ATP binding and can potentially influence binding of inhibitors to the ATP-binding site, MgCl2 was added.
the ED for the complete binding site can be added to the visualization (in cyan) if necessary.
An edge f→g is added to the graph only when at least one binding site is predicted around the target gene, so when sf,g≥1.
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