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Bound complexes were measured after pull down with magnetic protein A beads.
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The structures for the inhibitor bound complexes were retrieved from Protein Data Bank with PDB codes: 1AJX [32] for AHA001 bound complex, 1HVR [33] for XK263 bound complex and 1HXW [34] for ABT538 bound complex.
Free and bound complexes are resolved on 1.3% agarose gel.
The fraction of [S]-Met-tRNAi bound to 40S·eIF1·eIF1A or 40S·eIF1·eIF1A·mRNA complexes was measured using a PhosphorImager, plotted against the 40S subunit concentration, and the data were fit with a hyperbolic or quadratic binding equation, with the latter employed for tight binding.
A crystal structure of the inhibitor bound complex is reported.
Each bound complex was simulated twice using different starting velocities.
The amount of aminoacylated tRNAs that can be bound to the A-site of the ribosome in a ternary complex was measured using [H]Phe-tRNAPhe in poly(U -dependent assays.
The ability of DNA to bind the epitope and impede the formation of the epitope- antibody complex was measured in competition ELISAs (Fig. 4A).
Binding affinities of IL-4RαECD to the complex of IL-13/IL-13variant IL-13/IL-13variant IL-13/IL-13varianthe COINJECT command.
Binding affinities were measured by ITC.
After precipitation of the immune complexes, the amount of bound labeled insulin was measured with a liquid scintillation detector (1450 MicroBeta Trilux; Perkin Elmer Life Sciences, Turku, Finland), and the results were given as counts per minute.
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