To determine the generation of reactive oxygen species (ROS), we used a modified assay for the intracellular conversion of nitro blue tetrazolium (NBT) to formazan by superoxide anion.
If modifications to the SGF recipe are to be adopted, we suggest that they not only should be theoretically appealing but also should be accompanied by empirical data supporting the improved relevance of the modified assay to allergenicity assessment.
For the binary interaction assays, a modified assay buffer composed of 25 mM Hepes-KOH pH 7.2, 25 mM Tris HCl, 125 mM potassium acetate, 5 mM magnesium acetate, 2 mM EDTA, 2 mM EGTA, 10 mM dithiothreitol, 0.2% Igepal CA-630 (Nonidet P40 substitute) and 0.1 mg/ml BSA was used.
To test the role of specific versus unspecific adhesion, we monitored stable-bleb cell migration in various adhesive and non-adhesive environments using a modified assay that consists of two parallel glass slides coated with either Fibronectin, poly-L-lysine (PLL), BSA, or polyethyleneglycol (PEG) separated by 15-μm beads.
Serial dilution of samples indicates that the modified assay displays a linearity of 0.98 and an assay sensitivity (least detectable dose) of 1.3854 nmol/l.
The sensitivity and specificity rates of the modified assay in comparison with those of conventional culture were respectively 94.3%and99.2%2%.
We examined habituation in the lines showing chaining or high levels of male male courtship in a modified assay in which pairs of GAL4/ UAS-fru MIR males from each line were placed together and the amount of courtship measured both immediately after they were placed together and again after they had been together for 1 hr.
Endogenous Sirt1 activity in muscle cells and mouse white adipose tissue was measured using a modified assay with 5 μL of cell lysate.
The modified assay was used for identification of various genotypes of the HBB gene which were confirmed by direct sequencing.
The key features and performance descriptors of this modified assay are summarized in Table 2. Overall, the CV value for individual wells of Tx-Xyn-AF7 control varied between 8% to 11%, indicating that this screen was sufficiently reliable for library screening.
