After removing excess antibody, streptavidin-peroxidase was added, which binds to the secondary biotinylated antibody.
During phosphate starvation, the CDK inhibitor Pho81 binds to the secondary metabolite myo-D-inositol heptakisphosphate (IP7) and inhibits the Pho85-Pho80 complex [ 16, 17].
We have previously shown that 100 nM BODIPY-TMR-CGP binds to the secondary low affinity conformation of the β1-adrenoceptor, and as such may cause allosteric effects that result in an enhanced dissociation rate of 100 nM BODIPY-TMR-CGP (i.e., negative cooperativity) from the primary high-affinity β1-adrenoceptor conformation.
Relative spatial arrangement of the two binding sites suggests that the dsDNA molecule bound to the secondary DNA-binding site can sterically hinder the access of ssDNA into the primary ssDNA-binding site (Kagawa et al, 2008).
This was reflected in a significant increase in the fraction of BODIPY-TMR-CGP bound to the secondary, low-affinity site with increasing concentration of BODIPY-TMR-CGP from 10 nM (12.9 ± 1.1%, n = 3) to 100 nM (42.8 ± 0.9%, n = 3) respectively (P < 0.05, two-way anova followed by Bonferroni's post hoc test; Table 2005).
ppGpp is known as a transcriptional regulator [48], [49], and DksA, which binds to the RNA polymerase secondary channel, potentiates the effects of (p ppGpp on transcription.
It could be bound to the biotinylated secondary antibody of sandwich immunocomplex by biotin streptavidin conjugation to catalyze a CL reaction on a protein array, which produced strong CL emission.
An important mechanism of gene regulation, alternative splicing, is a complex process requiring over 100 splicing factors that recognise regulatory elements such as exonic and intronic splicing enhancers and silencers (ESEs, ESSs, ISEs, ISSs), which bind trans-acting splicing regulatory factors [ 13] that bind to the mRNA secondary structure [ 14].
Simberg and co-workers introduced a model for the protein corona which includes "primary binders" that interact with the NPs surface at first followed by "secondary binders" that binds to the primary binders by way of protein protein interactions [39].
This poly(A) tail promotes degradation by the degradosome, which contains two RNA-degrading enzymes: polynucleotide phosphorylase and RNase E. Polynucleotide phosphorylase binds to the 3' end of RNAs and the 3' extension provided by the poly(A) tail allows it to bind to the RNAs whose secondary structure would otherwise block the 3' end.
After incubation and washing, strong magnets were used to remove all beads that bound to the primary or secondary antibody.
