This approach has been successfully applied to bacterial sulfotransferases, generating kinetic parameters that are in excellent agreement with those obtained using a radiolabel transfer assay (30, 31).
The phosphorylation for polymer/DNA complexes was determined with a radiolabel assay using [γ-32P]ATP.
The phosphorylation reaction of the polymers or the polymer/DNA complexes was determined with a radiolabel assay using [γ-32P]ATP.
(1) Clarke et al. developed a radiolabeling assay to track the attachment of small biomolecules to QD surfaces.
MurG was assayed using a coupled radiochemical assay, as described by Zawadzke et al., in which lipid intermediate I is generated in situ by membranes containing translocase MraY, and is then converted in the presence of [H]-UDPGlcNAc and MurG to give radiolabelled lipid intermediate II, which is extracted into n-butanol and quantitated by scintillation counting.
Creatinine was assayed using a spectrophotometric assay.
To more accurately profile the repeat addition processivity of the reconstituted enzymes, we assayed telomerase activity using a primer radiolabeled at its 5′ end rather than by extension with radiolabeled dNTPs.
Binding affinities were also estimated using a competitive displacement radiolabel receptor assay as previously described (Keyt et al, 1996) but substituting VEGFR2 ΔKD m for the KDR-IgG fusion protein, and using the fusion proteins secreted from insect cells.
IC50, Ki, and n H values were determined using a glucocorticoid receptor radiolabeled competition binding assay as described in "Materials and methods" section.
To test if Rab6A interferes with KIF1C motor domain binding to microtubules, in vitro transcribed and translated, radiolabelled, full-length KIF1C was tested for microtubule binding in the presence and absence of purified Rab6A using a microtubule co-sedimentation assay.
