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A CREB reporter assay and [S]-GTPγS binding were employed to assess the constitutive activity and the activation by UDP, UDP-glucose and -galactose and the cysteinyl leukotrienes LTC4 and LTD4.
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Chromatin immunoprecipitation (ChIP) using an antibody to the p65 RelA DNA binding, was employed to measure NF-κB activation and binding in the absence and presence of IKK2 inhibitor.
Next, their binding properties were employed to change the localization of the respective fluorescent proteins within cells.
To enhance the likelihood of identifying the GTP-binding proteins, two assays were employed to monitor GTP-binding activities: (i) guanosine 5'- 3-O-[35S]thio triphosphate 5'- 3-O-[35S]thio triphosphate 5'- 3-O-[35S]thio triphosphate 5'- 3-O-[35S]thio triphosphatesodium dodecyl sulfate-polyacrylamide GTP electrophoresis and electroblottingammao nitrocelluloS -bindingeS -binding
UV visible, FT-Raman and circular dichroism spectroscopic techniques were employed to determine the binding mode, binding constant, sequence specificity and conformational effects of amsacrine binding to native calf thymus DNA.
Despite the fact that several different methods were employed to score binding of the target protein to the beads, of the ≈400 hits originally isolated, only two proved to be ligands for TNF-α and one of these was a nonselective binder.
Heterogeneous binding models (bi-Langmuir and Freundlich isotherms) that yield information on binding sites affinity distribution and heterogeneity index were employed to characterise this process.
Furthermore, MM-PBSA calculations were employed to evaluate the binding free energy of the complex (Table S4, ESI†).
UV Visible spectral studies were employed to study the binding of Ag and Au NPs with Calf-thymus DNA (CT-DNA).
To further improve detection, other functionalities were employed to increase analyte binding [ 6].
Alexafluor 594 or 488 conjugated secondary (Invitrogen, Carlsbad, CA, USA) antibodies were employed to detect antigen binding.
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