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SAM binding by Ash2L was UV-dependent and could be competed off by excess amount of unlabeled SAM (1000 fold, ∼0.35 mM) (Fig. 3A).

Binding to the wildtype probe could be competed off by the addition of a 100-fold molar excess of a non-radiolabelled wildtype probe to the nuclear extract (lane 3) while the addition of an excess of mutated non-radiolabelled probe had no effect on binding activity (lane 4) confirming that binding to the −216 Ets site was specific.

The nuclear extracts from either ARPE-19 cells or from FHL124 cells showed a strong binding to CREB site (lane 1, 6 of Fig. 4B), this binding can be competed off by the cold CREB oligo (lane 2, 7 of Fig. 4B) but not by the mutant oligo (lane 3, 8 of Fig. 4B).

Furthermore, it binds specifically to MD-MBA-435, MD-MBA-231, and HUVEC and can be competed off by free VCN.

7) Is it possible that eS26 interaction with Kaps is nonspecific and electrostatic, such that it can be competed off by Tsr2, a true binding partner?

We found that a labelled consensus sequence oligonucleotide bound too tightly to be competed off by the weaker competitor oligonucleotides at concentrations that could be feasibly achieved in the experiment.

The binding of iRGD-MBs on the endothelial cells was competed off by pre-incubation with the anti-αv or anti-β3 antibody (p < 0.01).

The binding assay showed that the binding of FITC-SERPINA3K to rMC-1 cells was competed off by excess concentrations of unlabeled SERPINA3K, but not by BSA (Fig. 7B), suggesting that the binding of SERPINA3K to Müller-derived rMC-1 cells is reversible and specific.

We assessed whether other polysome-associated proteins, many of which may have a role in translational regulation, can also be competed off polysomes by kcRNA.

Although Sgt1 bound in the low micromolar range (KD=1.64 μM), Cul1 bound Skp1 ∼70-fold more tightly, with a dissociation constant of ∼24 nM, so that Cul1 could not be competed off a Cul1 Skp1 complex by comparable levels of Sgt1.

This interaction could be competed off with unlabelled oligo and supershifted using the YB-1 antibody.