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The beads are magnetically captured, electrically stimulated and the light emission is proportional to the binding of ligand. A common label is Ruthenium (II) tri-bipyridine, NHS ester. electrodynamic forces London-Van der Waals forces: see DLVO theory. electrofocusing Any technique whereby chemical...
The canonical structure and function of centrioles are essential throughout mammalian development and physiology to form the cells’ cilia57. Centrioles serve as both the seed and template for cilia, and therefore, their precise structure is critical for their function57,58. Considering this major...
After washed at least three times with the lysis buffer, the agarose beads were resuspended in sodium dodecyl sulfate (SDS) sample buffer (Nacalai Tesque, 09499-14) and heated for 5 min at 95 °C. The supernatant was collected after centrifugation at 1000 g for 2 min. Immuno...
(or buffer-treated) beads were resuspended in buffer with or without 0.2% BSA. Either 300 ng of K18 fibrils or 30 ng of P301S recombinant tau were added to the bead suspension before mixing head-over-end for 30 min at room temperature. Following separation of the magnetic beads, ...
Beads elution was carried out by adding 200 mM glycine pH 2.5 to the beads and followed by the addition of 10% volume of neutralizing buffer (1M Tris, pH 10.4). The solution was then dialyzed against PBS by using Slide-A-Lyzer™ Dialysis Cassettes (Thermo Fisher Scientific, 10kDa). ...
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A small amount of 1 mm glass beads can be added and the mixture sonicated 3 times for about 5 seconds each time with a microprobe sonicator. The mixture is centrifuged and the protein concentration of the supernatant determined. One pg of protein from the soluble fraction of the culture can...
Bio-beads were removed and the nanodisc reconstitution mixture was bound again to Anti-FLAG resin (Biolegend) preequilibrated with 20 mM HEPES (pH 7.5), 150 mM NaCl at 4 °C for 2 h to remove free nanodiscs. The protein was eluted with 100 μg/mL FLAG peptide. The eluted ...
V.V. and J.T.G. acknowledge the support of the collaborative NRF CRP11-2012-02 grant, and MBI seed funds. A.R. and C.B. acknowledge financial support from seed founds of the Pontificia Universidad Católica de Chile. R.D. acknowledges funding from partly the National Medical Research Co...