200 µl lysate was incubated with GST-VAMPs, GST alone, syntaxin16-PrA or PrA alone bound to the appropriate beads in a volume of 1 ml at 4°C with rotation.
Recognizing that the optical mass of the bound molecules is responsible for inducing the observed LSPR shifts, the optical mass increase induced by bound analyte is defined as the product VD·ΔRI, where VD is the volume of analyte bound to the nanorod and ΔRI is the difference between the RI of the analyte and that of the surrounding medium.
HNE was diluted in 100 μL of PBS to a concentration of 1.0 mM and added to the Hsp90 protein bound to the Neutravidin resin, which gave a final volume of 400 μL and a final HNE concentration of 250 μM.
The proteases bound to the column were eluted with one bed volume of 0.9 M NaCl and dialysed at 4°C for 24 h in 1 l of 10 mM imidazole-buffer, pH 6.9.
The protein bound to the Sepharose was washed with 5 column volumes of Buffer 1.
To deplete Xenopus egg extracts of FANCJ, one volume of protein A-sepharose Fast Flow (PAS) (GE Healthcare, Piscataway, NJ, USA) was bound to 3 volumes of α- xlFANCJ serum or pre-immune serum through an overnight incubation at 4°C.
A natural dual to Lemma 3.2 is also given in [3], which provided a sharp lower bound for the volume of the polar of the Wulff shape W ν, f.
In [3], Schuster and Weberndorfer established a sharp bound for the volume of the Wulff shape W ν, f determined by an f-centered isotropic measure ν as follows.
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The contact stresses engendered are highly localised and bound to a very small volume of the material.
