Probes were labeled using 10X PCR labeling mix (Boehringer Mannheim, Biochemical Products, Indianapolis IN) at a concentration of 1X replacing the normal dNTP mix.
The relative potencies, expressed in T4 or T3 relative effect potency (REP) values, were either extracted from the publications or calculated by dividing the IC50 of the natural ligand T4 by the IC50 of the competitor, with IC50 as the concentration that is capable of replacing 50%% of a labeled TTR ligand from TTR.
The data were then fit to the equation 6with Keff given by eq 5 but with the concentration of TBA replaced by that of fatty acid, and with Fmax and Fmin being the fluorescent values for fully KcsA-bound and non-KcsA-bound Dauda, respectively.
We have investigated the role of water concentration by replacing various amounts of the water in enzymatic hydrolysis with oleyl alcohol, an inert oil that does not directly affect the function of the enzymes [ 38, 39].
Runs with 14% WIS were performed on whole pretreated slurry (SSF 2A to 2D in Table 1), on slurry with 20% lower inhibitor concentration (obtained by replacing part of the hydrolysate with water) (SSF 3A to 3D in Table 1) and on washed slurry (SSF 4A to 4C in Table 1) (obtained by diluting and washing with excess water several times).
Then, culture plates were placed in a 37°C, 5% CO2 humidified incubator for 21 days, and medium containing indicated concentration of nicotine was replaced once at the beginning of the week.
Lead has high affinity for calcium-binding sites in the proteins; picomolar concentration of lead can replace calcium in micromolar concentrations.
For example, in limb development the creation and maintenance of a concentration gradient of FGFs is crucial for the function of the apical ectodermal ridge in promoting limb outgrowth, since only a focal concentration of FGFs can replace the apical ectodermal ridge, whereas exposure of the entire embryo to FGFs will not result in additional limb development [ 17].
Challenges to domoic acid were conducted with a similar 96-well plate setup, with domoic acid concentrations replacing those of the PTZ.
After an overnight adherence phase, TNF-α was added to the respective wells in a final concentration of 10 ng/mL by replacing 100 μL medium per well, whereas control cells received equal amounts of fresh medium.
