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TP radioligand binding assays were carried out at 30 °C for 30 min in 100 μl reactions in the presence of 0 40 nM [H] SQ29,548 for Scatchard analysis or in the presence of 20 nM [H] SQ29,548 for saturation radioligand binding experiments as previously described [10].
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Binding experiments were completed as previously described but WPBS washes 3 and 4 were omitted as minimal proteins were removed in the previous experiments with these washes.
In vitro binding experiments were performed as previously described [9] with 20-μm brain cryosections from transgenic APP23 (N = 2), Tg2576 (N = 2) and APPswe-PS1dE9 (N = 2) mice and wild-type control mice (N = 5) that were sacrificed after in vivo studies were finalized (Table 2).
Ribosome binding experiments were performed as previously described [24].
RNA binding experiments was measured as previously described [21], [22].
Saturation binding experiments were performed as previously described [ 28].
Competition binding experiments were performed as previously described.
Lipid-binding floatation experiments as well as the lipid transport assay of NBD-PA by Ups1-Mdm35 were performed as previously described [ 6, 11].
Serum samples were diluted, polyclonal IgG was captured, and binding experiments were carried out as previously described [31], [32].
Protein purification and binding experiments were carried out as previously described (Hood et al., 2013).
The expression, isolation, and purification of wild-type and drug-resistant HIV-1 protease variants used for crystallization and binding experiments were carried out as previously described.
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