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The effect of radioiodination of minibody on PSCA binding was evaluated using SKW6.4 and SKW6.4-PSCA cells.
Binding was evaluated using flow cytometry.
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The metal binding capability was evaluated using the binding energy.
Furthermore, the physical binding environment of the binding pocket was evaluated using SiteMap [43, 44].
The NF-κB binding activity was evaluated using a LightShift Chemiluminescent EMSA kit (Pierce, Rockford, IL, USA).
In addition, the ability of the VvMYB5bR and VvMYB5bL proteins to bind MBS (MYB binding sites) cis-elements was evaluated using EMSA (Electrophoretic Mobility Shift Assay).
Their binding affinity with MDM2 was evaluated using both fluorescence polarization (FP) assay and 2D-NMR-HSQC experiments.
Capillary isoelectrofocusing (cIEF) and cation exchange chromatography (CEX) were used to evaluate charge heterogeneity; glycosylation profiles were assessed through hydrophilic interaction liquid chromatography (HILIC); aggregates content was evaluated through size exclusion chromatography (SEC) while binding affinity to FcRn was evaluated using isothermal titration calorimetry (ITC).
The binding affinity of FPyPEGCBT-c RGDfK) was evaluated using a standard in vitro binding assay on U-87 MG and SKOV-3 cancer cells expressing αVβ3, αVβ5 and α5β1 integrins and compared to a reFPyPEGCBT-c RGDfK(RGDfV) (Fig. 2).
In vitro integrin-binding of FPyPEGCBT-c RGDfK) was evaluated using a standard displacement assay oFPyPEGCBT-c RGDfKn.
The direct binding of each peptide to SIP1 was evaluated using the dot-blot binding assay as described elsewhere [37].
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