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This review will focus on established and newly emerging strategies for identifying and characterizing enzyme intermediates using a rapid transient kinetic approach.
This review focuses on the established and newly emerging strategies for identifying and characterizing enzyme intermediates using a rapid transient kinetic approach.
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To determine the binding affinity of WT and mutant enzymes to the dNTP substrate (KD-dNTP) and to estimate the maximum rate of dNTP incorporation by these enzymes (kpol), we carried out transient-state experiments using a rapid quench instrument (RQF-3, Kintek Corporation, Clarence, PA) at 37°C in RT buffer (50 mM Tris-HCl, pH 7.8 and 50 mM NaCl).
To test the function of cacao TcLEC2 in regulation of gene expression and to identify its putative downstream targets, a rapid transient transformation assay using cacao leaf tissue was utilized [ 38] (see Methods, Additional file 3).
Using immunoblotting and phospho-specific antibodies, we found that E2 produces a rapid, transient phosphorylation of PKC (Figure 9A), AKT (Figure 9B), c-RAF (Figure 9C), ERK1/2 (Figure 9D) and CREB (Figure 9E).
Our findings suggest a rapid, transient induction upon DEE exposure.
ET-1 (100 nM) stimulated a rapid transient increase in expression of Atf3 mRNA in cardiomyocytes.
This is consistent with a rapid, transient effect of psychological stress on sickness responses to typhoid vaccine in humans.
A rapid transient increase in cytosolic Ca2+ levels occurs when E. purpurea extracts are applied to HEK293 cells.
Moreover, a rapid transient up-regulation of CSN3 is observed in response to various growth inhibition stimuli [ 3].
We noticed that addition of glucose to derepressed reg1Δ cells led to a rapid, transient increase in SUC2 expression.
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