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Binding was further characterized by surface plasmon resonance (SPR), allowing determination of binding affinities and kinetic constants.
Fundamental to any thermodynamic characterization of the drug DNA interactions is the determination of binding constants.
Open image in new window Figure 3 Determination of binding affinities between hENT1 and adenosine, deoxyadenosine, or adenine.
They obtained highly stable signals that enabled multiple measurements with a single nanopipette, thereby allowing the determination of binding constants.
For the determination of binding affinities, dissociation constants (K d ) were estimated by gel shift assay using SYBR Gold.
In vitro determination of binding and release kinetics of TG-PDGF.AB allowed proof of concept of the developed binding technology.
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Precise determinations of binding site locations are also enabled by PAR-CLIP [ 31].
The proportionality constant that relates this occupancy score to the true occupancy of the promoter region sequence is unknown but is not needed because we use a correlation coefficient to compare the occupancy score to the experimental determinations of binding locations.
The binding sites of Spo0J determined using this technique were consistent with those reported earlier, clearly indicating that the ChAP-chip methods are sufficiently sensitive for the genome-wide determination of specific binding sites of DNA-binding proteins.
The ability to quantify the strength with which two binding partners interact is desirable but the accurate determination of equilibrium binding constants is a difficult process.
Determination of the binding site of RNA [poly(U ] reveals that the binding sites for poly(U) and ATP do not overlap (see Figures S1 S3 of the Supporting Information).
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