In brief, FDP concentration was measured by an enzymatic procedure of Racker (Bergmeyer 2012) using a commercial JSAY139 FDP kit (Jie Shi Kang Biotech Co., Ltd. China).
A brief, high concentration of carbachol, to cause SR Ca2+ depletion without eliciting receptor-operated channel opening, also produced these results.
In brief, protein concentration was determined by Bradford assay [ 75] and total protein resolved by SDS-PAGE followed by transfer to nitrocellulose membrane.
In brief, RNA concentration was determined by absorbency at 260 nm, and quality and integrity was verified using the RNA 6000 Nano assay on the Agilent 2100 Bioanalyzer (Agilent Technologies).
The profile of the rhodamine visualization of the picospritz event (Fig. 1D and Supplementary movie 1) indicates a very brief glutamate concentration peak (a few tens of milliseconds), while the neuronal response clearly extends for a much longer period, far longer than synaptic events in these cells (Kirby et al., 2009).
In brief, different concentrations of test compound were prepared by serial dilution and 20 μl of each concentration was added to the rows of a 96-well microtitre plate in diminishing concentrations.
In brief, various concentrations of the competitor peptides were incubated for 4 hours at 37°C, with constant concentrations of biotinylated hemagglutinin (HA) (307 319) peptide (0.5 nM for DR1, 5 nM for DR4) and DR molecule (10 nM) in PBS containing OcG.
In brief, different concentrations of prodomain constructs (2 50 nM) were pre-incubated with 2 nM falcipain-2 in 100 mM sodium acetate, 5 mM DTT, pH 5.5 for 10 min at room temperature.
In brief, different concentrations of insulin (bovine pancreas, Sigma) ranging from 0.5 2 mg/mL (85 340 µM) were prepared in 25 mM HCl (pH 1.6) and placed at 60°C for an hour (protofibrils) or at least overnight (fibrils) although incubation times were dependent on protein concentration.
