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These biotinylated nucleic acid complexes were incubated with protein lysates and bound complexes were captured on streptavidin magnetic beads and analyzed by immunoblotting.
These biotinylated RNA-DNA hybrids were incubated with protein lysate isolated from transiently transfected 293T cells and bound complexes were captured on streptavidin-labeled magnetic beads using µMACS columns.
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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.
Immune complexes were captured on protein A Sepharose (Sigma, Poole, UK) and, after washing, the quantity of radiolabelled antigen bound was determined by scintillation counting.
DNA RNA complexes were captured using the Agilent Bravo instrument.
DNA/protein complexes were captured by Dynabeads Protein A (Invitrogen).
Free and bound complexes are resolved on 1.3% agarose gel.
A crystal structure of the inhibitor bound complex is reported.
Each bound complex was simulated twice using different starting velocities.
In short, the partition complex is captured by ParA-ATP dimers that are bound to DNA.
DNA bound to probes was captured with streptavidin-coated magnetic beads (Promega Corporation, Madison, Wisconsin, USA).
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