As shown in Fig. 6A, the increase in Cl anions to physiological conditions did not affect binding, and interference was only detected at a higher salt concentration, 350 mM, for both proteins.
The assay employs an incubation step at acidic pH, which was shown to decrease nonspecific binding and interference from IL-13 binding proteins.
Our observations suggest that these mutants restrict wild-type PPARγ action via a non-DNA binding, transcriptional interference mechanism, which may involve sequestration of functionally limiting coactivators.
The improvement of assay precision on endogenous IL-13 measurement at pH 3.5 may be linked to the reduction of binding proteins interference.
Like DCC, integrin β5 interferes microtubule binding, whereas the interference is relatively weaker than DCC.
We conclude that RelB regulates TNFα cytokine synthesis by competitive interference binding with RelA, which leads to downregulation of TNFα production.
Inhibition of Drp1 function using either expression of DrpK38A, a dominant negative mutant defective in GTP binding, or RNA interference, leads to the formation of a highly fused and tubular mitochondrial network, thus implicating Drp1 in mitochondrial fission [6], [8].
This inherent limitation is also true of other fundamental biological assays based on lysed cells, where the binding chemistries cause interference in the analyses.
In fact, earlier fluorescent biosensors utilizing pyrene-appended aptamers have shown perturbation of target binding due to interference of the binding site by dye-labeling in the binding pocket.
Oligonucleotides for plasminogen activator inhibitor 1 RNA binding protein (PAI RBP) interference were pre-designed and synthesised by Ambion Inc. (Austin, TX, USA).
